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The fission yeast Schizosaccharomyces octosporus is one of four species in the genus Schizosaccharomyces. Recently released genome sequence data provide useful information for comparative studies. However, Sz. octosporus has not yet been genetically characterized because there have been no heterothallic strains of this species. Here we report the construction of stable heterothallic strains of Sz. octosporus for genetic crosses. First, we continuously observed the mating process of a homothallic strain, yFS286, and determined the mating frequency of Sz. octosporus on various sporulation media. It showed, on average, 30% zygote formation on mating, and a higher frequency of zygotes (43.8 ± 4.7%) on PMG medium. Regardless of sporulation, the number of spores within an ascus was variable. Schizosaccharomyces octosporus forms eight-spored asci, but preferentially produced four-spored asci on MEA or YMoA medium. To obtain heterothallic strains essential for genetic analyses, we isolated spontaneous mutants showing heterothallic-like phenotypes. We also constructed stable heterothallic strains by deleting the silent mat region. As a result, we established the following heterothallic strains, TS162 as h+ and TS150/TS161 as h-, which successfully mated with each other. These genetic tools will be useful for yeast genetics such as molecular cloning, gene complementation tests and tetrad (octad) analysis., (© FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

A lightweight-compact variable-gap undulator (LCVGU) having the force-cancellation system based on the multipole monolithic magnets (MMMs) has been developed. The LCVGU is free from the heavy mechanical frames, which is a fundamental element specific to conventional variable-gap undulators (VGUs) because of a strong attractive force, and thus the cost and time for construction and installation are expected to be significantly reduced; the MMMs counteract the strong attractive force in a cost-effective manner. Results of mechanical tests and magnetic-field measurements of two prototype LCVGUs equipped with the proposed force cancellation system have revealed the comparable performance with the conventional VGUs.

A woman in her 70s was diagnosed with a protruding mucosa-associated lymphoid tissue (MALT) lymphoma during a secondary health examination. After eradication of Helicobacter pylori, a biopsy revealed gastric follicular lymphoma (FL) and the lesion was still protruding one year later. 18 F-fluorodeoxyglucose positron emission tomography showed focal nodular hypermetabolic activity, suggesting that FL may have transformed into a diffuse large B-cell lymphoma. Upper gastrointestinal endoscopy, colonoscopy, and capsule endoscopy showed no other lesions in the gastrointestinal tract, and bone marrow biopsy showed no permeation into the marrow. Therefore, this lesion, which appeared as a submucosal tumor, was limited to the stomach. Laparoscopy and endoscopy cooperative surgery was performed, because it allows for correct pathological diagnosis while removing only a minimal portion of the stomach wall. Histological findings showed follicular structures consisting of abnormal lymphoid cells. Immunohistochemical analysis revealed that neoplastic cells were positive for CD20, CD79a, Bcl-2, CD10, and c-MYC, but negative for CD3, CD5, and cyclin D1. Finally, we diagnosed this lesion as a primary gastric FL.

Background: Recently, morbidities due to primary cytomegalovirus (CMV) infection have increased in young Japanese adults because of decreased anti-CMV antibodies in them. CMV infections are typically resolved naturally in immunocompetent individuals, and complications rarely occur. Here we present the case of an immunocompetent adult with CMV infection complicated by splenic infarctions and an esophageal ulcer., Case Report: A 37-year-old male complaining of a prolonged fever and liver injury was admitted to hospital for a closed examination. The patient had general malaise and mild appetite loss but no abdominal pain. Symptoms of infectious mononucleosis, including liver injury, appearance of atypical lymphocytes in the blood, and hepatosplenomegaly, were observed. A primary CMV infection was confirmed by CMV-IgM positive and CMV-IgG negative serological tests. Enhanced abdominal computed tomography confirmed hepatitis and splenic infarction, and an upper gastrointestinal endoscopy revealed an esophageal ulcer. The patient exhibited no predisposing risk factors for thrombosis, and he was diagnosed with splenic infarctions associated with CMV infection. Because the patient was immunocompetent, he underwent symptomatic therapy without antiviral or anticoagulant therapies. The treatment improved his overall condition. Including the present case, only 11 cases of CMV infections with splenic infarction in immunocompetent individuals have been reported. Contrary to what is observed in immunocompromised hosts, upper gastrointestinal lesions with CMV infection are rare in immunocompetent individuals. The esophageal lesion observed in our patient was a typical punched-out ulcer. The immunohistochemical staining of the tissue biopsies revealed that the ulcer was associated with CMV., Conclusion: Although splenic infarctions and esophageal ulcers are rare, they should be considered as potential complications accompanying CMV infection in immunocompetent individuals. The administration of symptomatic therapy should be considered even when the patient is immunocompetent.

A 40-year-old man complaining of abdominal distention was referred to our hospital. Computed tomography of the abdomen demonstrated a very large abdominal mass with fat and calcification. The size of the mass rapidly increased from 30cm to 40cm over two weeks. The tumor was removed and diagnosed by pathological examination to be a retroperitoneal mature cystic teratoma that contained a 40-cm long, mature intestinal tract-like cyst, together with bone marrow and fat. The rapid growth of the tumor may have been caused by an increased secretion in the cyst.

Background and study aims  It is important to examine the pharynx during upper gastrointestinal endoscopy. Pharyngeal anesthesia using topical lidocaine is generally used as pretreatment. In Japan, lidocaine viscous solution is the anesthetic of choice, but lidocaine spray is applied when the former is considered insufficient. However, the relationship between the extent of pharyngeal anesthesia and accuracy of observation is unclear. We compared the performance of lidocaine spray alone versus lidocaine spray combined with lidocaine viscous solution for pharyngeal observation during transoral endoscopy. Patients and methods  In this prospective, double-blinded, randomized clinical trial conducted between January and March 2015, 327 patients were randomly assigned to lidocaine spray alone (spray group, n = 157) or a combination of spray and viscous solution (combination group, n = 170). We compared the number of pharyngeal observable sites (non-inferiority test), pain by visual analogue scale, observation time, and the number of gag reflexes between the two groups. Results  The mean number of images of suitable quality taken at the observable pharyngeal sites in the spray group was 8.33 (95 % confidence interval [CI]: 7.94 - 8.72) per patient, and 8.77 (95 % CI: 8.49 - 9.05) per patient in the combination group. The difference in the number of observable pharyngeal sites was - 0.44 (95 % CI: - 0.84 to - 0.03, P  = 0.01). There were no differences in pain, observation time, or number of gag reflexes between the 2 groups. Subgroup analysis of the presence of sedation revealed no differences between the two groups for the number of pharyngeal observation sites and the number of gag reflexes. However, the number of gag reflexes was higher in the spray group compared to the combination group in a subgroup analysis that looked at the absence of sedation. Conclusions  Lidocaine spray for pharyngeal anesthesia was not inferior to lidocaine spray and viscous solution in terms of pharyngeal observation. It was considered that lidocaine viscous solution was unnecessary for pharyngeal observation. UMIN000016073.

We report an unexpectedly large flux loss observed in permanent magnets in one of the undulators operated in SACLA, the x-ray free electron laser facility in Japan. Characterizations of individual magnets extracted from the relevant undulator have revealed that the flux loss was caused by a homogeneous magnetization reversal extending over a wide area, but not by demagnetization of individual magnets damaged by radiation. We show that the estimated flux-loss rate is much higher than what is reported in previous papers, and its distribution is much more localized to the upstream side. Results of numerical and experimental studies carried out to validate the magnetization reversal and quantify the flux loss are presented, together with possible countermeasures against rapid degradation of the undulator performance.

The diversification of sex pheromones is regarded as one of the causes of prezygotic isolation that results in speciation. In the fission yeast Schizosaccharomyces pombe, the molecular recognition of a peptide pheromone by its receptor plays an essential role in sexual reproduction. We considered that molecular coevolution of a peptide-mating pheromone, M factor, and its receptor, Map3, might be realized by experimentally diversifying these proteins. Here, we report the successful creation of novel mating-type pairs by searching for map3 suppressor mutations that rescued the sterility of M-factor mutants that were previously isolated. Several strong suppressors were found to also recognize WT M factor. The substituted residues of these Map3 suppressors were mapped to F204, F214, and E249, which are likely to be critical residues for M-factor recognition. These critical residues were systematically substituted with each of the other amino acids by in vitro mutagenesis. Ultimately, we successfully obtained three novel mating-type pairs constituting reproductive groups. These novel mating-type pairs could not conjugate with WT maters. Furthermore, no flow of chromosomally integrated drug-resistance genes occurred between the novel and the WT mating pairs, showing that each experimentally created reproductive group [e.g., M factor(V5H) and Map3(F214H)] was isolated from the WT group. In conclusion, we have succeeded in creating an artificial reproductive group that is isolated from the WT group. In keeping with the biological concept of species, the artificial reproductive group is a new species.

Background: Recurrence and complication rates of pressure sores are highest in the ischial region, and other donor sites are needed for recurrent pressure sores. The potential of a new freestyle pedicle flap for ischial lesions, an internal pudendal artery perforator (iPap) thigh flap, was examined through anatomical and theoretical analyses and a case series using computed tomography angiography., Methods: The skin flap was designed in the thigh region based on an iPap. The skin perforators were marked with a Doppler probe. One patient underwent computed tomography angiography with fistulography to identify the damage to or effects on the pedicle vessels of the flap. Debridement of ischial lesions and flap elevation were performed in the jackknife position., Results: The iPap thigh flaps were performed in 5 patients, 4 with ischial pressure sores and 1 with calcinosis cutis of the ischial region. The width and length of the flaps ranged from 5 to 8 cm (mean, 6.6 cm) and 10 to 17 cm (mean, 12.6 cm), respectively. Three patients underwent partial osteotomy of the ischial bone. No complications, including flap necrosis or wound dehiscence of the donor and reconstructed sites, were observed., Conclusions: The perforator vessels of the internal pudendal artery are very close to the ischial tuberosity. Blood flow to the flap is reliable when careful debridement of the pressure sore is performed. The iPap thigh flap is a new option for soft-tissue defects in the ischial region, including ischial pressure sores.

Background: When a tumor in the lateral eyebrow region is resected, reconstruction of the eyebrow and upper eyelid defects is necessary. We perform simultaneous reconstruction of such defects; sideburn hair is included on an extended lateral orbital flap. We describe our method and results of a retrospective evaluation., Methods: We treated 6 patients with partial eyebrow and upper eyelid defects. An extended lateral orbital flap was designed on the lateral region of the lateral canthus including sideburn hair and was elevated with a pedicle of the orbicularis oculi muscle. Flap size, surgical outcomes, and reconstructed eyebrow morphology were evaluated., Results: Mean flap size was 2.7 × 4.1 cm, with a maximum width and length of 3.5 and 5.1 cm, respectively. The mean hair-bearing region was 1.4 × 0.9 cm, with a maximum width and length of 1.8 and 1.3 cm, respectively. Total flap survival and hair growth were confirmed in all patients. The reconstructed eyebrow and eyelid were aesthetically satisfactory. The new eyebrow hair grew long and was directed upward but considered acceptable. No additional surgery was performed for any patient. The donor site scar was acceptable because it followed the wrinkles of the lateral eyelid. No paralysis of the temporal branch of the facial nerve resulted., Conclusions: The procedure for raising an extended hair-bearing lateral orbital flap is relatively easy, although attention must be paid to the temporal facial nerve. This flap is useful for simultaneously reconstructing defects of the upper eyelid and lateral eyebrow.

Background: Cutaneous malignant melanoma has a poor prognosis. The detrimental effect of incisional biopsies on the outcome of malignant melanoma has been debated. The aim of this study was to determine the effect of the presence and type of biopsy on the prognosis of malignant melanoma., Methods: The medical records of 109 malignant melanoma patients treated at Tokushima University Hospital from 1983 to 2007 were reviewed. After excluding 28 cases with stage 0 disease or incomplete data, 81 cases were analyzed in detail with respect to patient sex, age, tumor site, clinical stage at diagnosis, presence of ulceration or lymph node metastasis, and prognosis. The five-year survival and five-year disease-free survival rates of patients who underwent incisional or excisional biopsies were compared with those who did not undergo a biopsy., Results: The male-to-female ratio was 1:1.19. The mean age was 61.3 years (range, 19-93 years). The most common site was a lower extremity, and the most common clinical stage was stage II. No significant differences in clinicopathological features, five-year survival rates, and five-year disease-free survival rates were observed among the three groups., Conclusions: The presence and type of biopsy neither affected the metastatic rate nor the prognosis of malignant melanoma. The use of incisional biopsies is not encouraged because tumor thickness cannot be measured accurately. However, they may be helpful for confirming the diagnosis if an excisional biopsy cannot be performed.

A new soft X-ray beamline, BL07LSU, has been constructed at SPring-8 to perform advanced soft X-ray spectroscopy for materials science. The beamline is designed to achieve high energy resolution (E/ΔE> 10000) and high photon flux [>10(12) photons s(-1) (0.01% bandwidth)(-1)] in the photon energy range 250-2000 eV with controllable polarization. To realise this state-of-the-art performance, a novel segmented cross undulator was developed and adopted as a light source. The details of the undulator light source and beamline monochromator design are described. The achieved performance of the beamline, such as the photon flux, energy resolution and the state of polarization, is reported.

The newly installed BL28XU beamline at SPring-8 is dedicated to in situ structural and electronic analysis of rechargeable batteries. It supports the time range (1 ms to 100 s) and spatial range (1 µm to 1 mm) needed for battery analysis. Electrochemical apparatus for battery charging and discharging are available in experimental hutches and in a preparation room. Battery analysis can be carried out efficiently and effectively using X-ray diffraction, X-ray absorption fine-structure analysis and hard X-ray photoelectron spectroscopy. Here, the design and performance of the beamline are described, and preliminary results are presented.

The aim of this retrospective study was to examine whether serum albumin levels offer a good marker of nutritional status in patients with burns. Serum albumin levels have been used to evaluate nutritional status in burns patients, even though these levels are affected by various factors and are not specific to malnutrition. To clarify whether provision of nutrition or presence of inflammation affects serum albumin levels, we studied serum albumin levels, C-reactive protein (CRP) levels and caloric intake over time in 30 patients with burns. Serum albumin levels did not respond to provision of nutrition, but correlated negatively with CRP levels, suggesting that serum albumin levels are more closely associated with inflammation than nutrition. This study also suggests that hypoalbuminemia is a good indicator of the severity of burns or associated complications. We conclude that serum albumin levels do not offer a good nutritional marker in burns patients.

Mating pheromone signaling is essential for conjugation between haploid cells of P-type (P-cells) and haploid cells of M-type (M-cells) in Schizosaccharomyces pombe. A peptide pheromone, M-factor, produced by M-cells is recognized by the receptor of P-cells. An M-factor-less mutant, in which the M-factor-encoding genes are deleted, is completely sterile. In liquid culture, sexual agglutination was not observed in the mutant, but it could be recovered by adding exogenous synthetic M-factor, which stimulated expression of the P-type-specific cell adhesion protein, Map4. Exogenous M-factor, however, failed to recover the cell fusion defect in the M-factor-less mutant. When M-factor-less cells were added to a mixture of wild-type P- and M-cells, marked cell aggregates were formed. Notably, M-factor-less mutant cells were also incorporated in these aggregates. In this mixed culture, P-cells conjugated preferentially with M-cells secreting M-factor, and rarely with M-factor-less M-cells. The kinetics of mating parameters in liquid culture revealed that polarized growth commenced from the contact region of opposite mating-type cells. Taken together, these findings indicate that M-factor at a low concentration induces adhesin expression, leading to initial cell-cell adhesion in a type of "distal pheromone action", but M-factor that is secreted directly in the proximity of the adhered P-cells may be necessary for cell fusion in a type of "proximal pheromone action".

Thrombin-activatable fibrinolysis inhibitor (TAFI, carboxypeptidase B2) is a 58-kDa plasma glycoprotein secreted by hepatocytes as an inactive form. TAFI is activated by the thrombin-thrombomodulin complex, and activated TAFI (TAFIa) plays an important role in regulating the balance between coagulation and fibrinolysis through inhibition of fibrinolysis. It has been suggested that high levels of TAFI in circulating plasma increase the risks of cardiovascular death and acute phase in ischaemic stroke. However, the mechanisms of regulating TAFI expression have been unclear. The present study investigated the effects of nobiletin (a polymethoxy flavonoid contained in the rind of citrus fruits) on TAFI gene (CPB2) and TAFI antigen expression in cultured human hepatoma HepG2 cells. Nobiletin decreased the release of TAFI antigen from HepG2 cells into conditioned medium in parallel with decreased levels of CPB2 mRNA and antigen. The half-life time of CPB2 mRNA in nobiletin-treated cells was unchanged compared to that of untreated control cells. Using nobiletin-treated cells that were transfected with a luciferase CPB2 promoter reporter plasmid, activity decreased to half of that in untreated control cells. A series of luciferase reporter constructs containing 5´-flanking region deletions of the human CPB2 gene showed that the sequences from -150 bp to -50 bp were essential for transcription of CPB2 and contained an AP-1 binding sequence at ~ -119 bp to - 99 bp in the CPB2 promoter. The amount of complexed nuclear protein and sequences from ~ -119 bp to -99 bp was decreased in nobiletin-treated cells. ChIP assays showed that c-Jun bound to the ~ -119 bp to -99 bp region of the CPB2 promoter and that the amount of the immunocomplex decreased after nobiletin treatment. Therefore, nobiletin-induced repression of CPB2 transcription might involve AP-1 inhibition and/or prevention of AP-1 binding in a specific region on the CPB2 gene in HepG2 cells.

The mating reaction is triggered by specific pheromones in a wide variety of organisms. Small peptides are used as mating pheromones in yeasts and fungi. In the fission yeast Schizosaccharomyces pombe, M-factor is a C terminally farnesylated nonapeptide secreted from M-cells, and its counterpart, P-factor, is a simple peptide composed of 23 amino acids. The primary structure requirements for the biological activity of pheromone peptides remain to be elucidated. Here, we conducted comprehensive substitution of each of the amino acids in M-factor peptide and inspected the mating ability of these missense mutants. Thirty-five sterile mutants were found among an array of 152 mutants with single amino acid substitutions. Mapping of the mutation sites clearly indicated that the sterile mutants were associated exclusively with four amino acid residues (VPYM) in the carboxyl-terminal half. In contrast, the substitution of four amino-terminal residues (YTPK) with any amino acid had no or only a slightly deleterious effect on mating. Furthermore, deletion of the three N-terminal residues caused no sterility, although truncation of a fourth residue had a marked effect. We conclude that a farnesylated hexapeptide (KVPYMC(Far)-OCH(3)) is the minimal M-factor that retains pheromone activity. At least 15 nonfunctional peptides were found to be secreted, suggesting that these mutant M-factor peptides are no longer recognized by the cognate receptor.

Background: Alveolar bone grafting is a standard procedure used to achieve good occlusion for both functional and aesthetic purposes in patients with cleft lip and palate. At the past, main methods used to evaluate bone bridge formation after bone grafting are radiographs, such as dental, occlusal, and panoramic., Purpose: To evaluate bone bridge both qualitatively and quantitatively, we used CT scans (conventional and QCT). Quantitative computed tomography (QCT) has previously been used for measuring bone mineral density of the lumbar vertebrae., Patients and Methods: The study comprised 26 male and 15 female patients who underwent alveolar bone grafting. We analyzed bone bridge with regard to four factors: marginal bone level, vertical height, anteroposterior bone width and bone mineral density using dental radiographs, and CT scans such as conventional and QCT. The clinical results of orthodontic treatment were evaluated more than 2 years postoperatively., Results: Orthodontic treatment was considered to be successful when the bone bridge satisfied the following criteria: marginal bone level >= 3, vertical height >= 6.5 mm, anteroposterior bone width >= 5 mm, and bone mineral density < 350 mg Ca(5) (PO(4)) OH/mL., Conclusion: we could predict the prognosis of patients' orthodontic treatment in early stage after bone grafting.

The incidence of brain metastasis is increasing, however, little is known about molecular mechanism responsible for lung cancer-derived brain metastasis and their development in the brain. In the present study, brain pathology was examined in an experimental model system of brain metastasis as well as in human brain with lung cancer metastasis. In an experimental model, after 3-6 weeks of intracardiac inoculation of human lung cancer-derived (HARA-B) cells in nude mice, wide range of brain metastases were observed. The brain sections showed significant increase in glial fibrillary acidic protein (GFAP)-positive astrocytes around metastatic lesions. To elucidate the role of astrocytes in lung cancer proliferation, the interaction between primary cultured mouse astrocytes and HARA-B cells was analyzed in vitro. Co-cultures and insert-cultures demonstrated that astrocytes were activated by tumor cell-oriented factors; macrophage migration inhibitory factor (MIF), interleukin-8 (IL-8) and plasminogen activator inhibitor-1 (PAI-1). Activated astrocytes produced interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interleukin-1 β (IL-1β), which in turn promoted tumor cell proliferation. Semi-quantitative immunocytochemistry showed that increased expression of receptors for IL-6 and its subunits gp130 on HARA-B cells. Receptors for TNF-α and IL-1β were also detected on HARA-B cells but down-regulated after co-culture with astrocytes. Insert-culture with astrocytes also stimulated the proliferation of other lung cancer-derived cell lines (PC-9, QG56, and EBC-1). These results suggest that tumor cells and astrocytes stimulate each other and these mutual relationships may be important to understand how lung cancer cells metastasize and develop in the brain.

Purpose: The heat shock protein 90 (Hsp90) plays an important role in chaperoning oncogenic client proteins in multiple myeloma (MM) cells, and several Hsp90 inhibitors have shown antitumor activities both in vitro and in vivo. However the precise mechanism of action of Hsp90 inhibitor in MM has not been fully elucidated., Experimental Design: We evaluated the antitumor activities of KW-2478, a nonansamycin Hsp90 inhibitor, in MM cells with various chromosomal translocations of immunoglobulin heavy chain (IgH) loci both in vitro and in vivo., Results: Our studies revealed that exposure of KW-2478 to MM cells resulted in growth inhibition and apoptosis, which were associated with degradation of well-known client proteins as well as a decrease in IgH translocation products (FGFR3, c-Maf, and cyclin D1), and FGFR3 was shown to be a new client protein of Hsp90 chaperon complex. In addition, KW-2478 depleted the Hsp90 client Cdk9, a transcriptional kinase, and the phosphorylated 4E-BP1, a translational inhibitor. Both inhibitory effects of KW-2478 on such transcriptional and translational pathways were shown to reduce c-Maf and cyclin D1 expression. In NCI-H929 s.c. inoculated model, KW-2478 showed a significant suppression of tumor growth and induced the degradation of client proteins in tumors. Furthermore, in a novel orthotopic MM model of i.v. inoculated OPM-2/green fluorescent protein, KW-2478 showed a significant reduction of both serum M protein and MM tumor burden in the bone marrow., Conclusions: These results suggest that targeting such diverse pathways by KW-2478 could be a promising strategy for the treatment of MM with various cytogenetic abnormalities., (Copyright (c) 2010 AACR.)

Following any type of brain injury such as lesion, stroke, and tumor/cancer invasion, microglia are rapidly activated and recruited to the site of injury. Microglia is the main immune effector cell population of the central nervous system and control immune cell recruitment. However, the molecular mechanism of brain metastasis and interaction between neuron-glia-tumor cells are poorly understood. Therefore, we established an animal model for brain metastasis using human lung cancer-derived cells (HARA-B) in nude mice. Accumulation of activated microglia was observed around tumor cells depending on the size of metastatic foci and the area of the brain. In vitro study, conditioned medium from primary cultured mouse microglia inhibited the proliferation of tumor cells, while tumor cell-conditioned medium inhibited the proliferation of primary cultured neurons from mouse cortex. Though the responsible factors released from microglia and tumor cells are still under investigation, these studies will contribute to understand the mechanism of cell-cell interaction in the brain and possible therapeutic target for brain metastasis.

It has been shown that molecular hydrogen (H(2)) acts as a therapeutic antioxidant and suppresses brain injury by buffering the effects of oxidative stress. Chronic oxidative stress causes neurodegenerative diseases such as Parkinson's disease (PD). Here, we show that drinking H(2)-containing water significantly reduced the loss of dopaminergic neurons in PD model mice using both acute and chronic administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The concentration-dependency of H(2) showed that H(2) as low as 0.08 ppm had almost the same effect as saturated H(2) water (1.5 ppm). MPTP-induced accumulation of cellular 8-oxoguanine (8-oxoG), a marker of DNA damage, and 4-hydroxynonenal (4-HNE), a marker of lipid peroxidation were significantly decreased in the nigro-striatal dopaminergic pathway in mice drinking H(2)-containing water, whereas production of superoxide (O(2)*(-)) detected by intravascular injection of dihydroethidium (DHE) was not reduced significantly. Our results indicated that low concentration of H(2) in drinking water can reduce oxidative stress in the brain. Thus, drinking H(2)-containing water may be useful in daily life to prevent or minimize the risk of life style-related oxidative stress and neurodegeneration.

Bradykinin (BK) has been reported to be a mediator of brain damage in acute insults. Receptors for BK have been identified on microglia, the pathologic sensors of the brain. Here, we report that BK attenuated lipopolysaccharide (LPS)-induced release of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta from microglial cells, thus acting as an anti-inflammatory mediator in the brain. This effect was mimicked by raising intracellular cAMP or stimulating the prostanoid receptors EP2 and EP4, while it was abolished by a cAMP antagonist, a prostanoid receptor antagonist, or by an inhibitor of the inducible cyclooxygenase (cyclooxygenase-2). BK also enhanced formation of prostaglandin E(2) and expression of microsomal prostaglandin E synthase. Expression of BK receptors and EP2/EP4 receptors were also enhanced. Using physiological techniques, we identified functional BK receptors not only in culture, but also in microglia from acute brain slices. BK reduced LPS-induced neuronal death in neuron-microglia co-cultures. This was probably mediated via microglia as it did not affect TNF-alpha-induced neuronal death in pure neuronal cultures. Our data imply that BK has anti-inflammatory and neuroprotective effects in the central nervous system by modulating microglial function.

CD38, a transmembrane glycoprotein with ADP-ribosyl cyclase activity, catalyses the formation of Ca2+ signalling molecules, but its role in the neuroendocrine system is unknown. Here we show that adult CD38 knockout (CD38-/-) female and male mice show marked defects in maternal nurturing and social behaviour, respectively, with higher locomotor activity. Consistently, the plasma level of oxytocin (OT), but not vasopressin, was strongly decreased in CD38-/- mice. Replacement of OT by subcutaneous injection or lentiviral-vector-mediated delivery of human CD38 in the hypothalamus rescued social memory and maternal care in CD38-/- mice. Depolarization-induced OT secretion and Ca2+ elevation in oxytocinergic neurohypophysial axon terminals were disrupted in CD38-/- mice; this was mimicked by CD38 metabolite antagonists in CD38+/+ mice. These results reveal that CD38 has a key role in neuropeptide release, thereby critically regulating maternal and social behaviours, and may be an element in neurodevelopmental disorders.

The IAN (immune-associated nucleotide-binding protein) family is a family of functionally uncharacterized GTP-binding proteins expressed in vertebrate immune cells and in plant cells during antibacterial responses. Here we show that all eight IAN family genes encoded in a single cluster of mouse genome are predominantly expressed in lymphocytes, and that the expression of IAN1, IAN4, and IAN5 is significantly elevated upon thymic selection of T lymphocytes. Gain-of-function experiments show that the premature overexpression of IAN1 kills immature thymocytes, whereas short hairpin RNA-mediated loss-of-function studies show that IAN4 supports positive selection. The knockdown of IAN5 perturbs the optimal generation of CD4/CD8 double-positive thymocytes and reduces the survival of mature T lymphocytes. We also show evidence suggesting that IAN4 and IAN5 are associated with anti-apoptotic proteins Bcl-2 and Bcl-xL, whereas IAN1 is associated with pro-apoptotic Bax. Thus, the IAN family is a novel family of T cell-receptor-responsive proteins that critically regulate thymic development and survival of T lymphocytes and that potentially exert regulatory functions through the association with Bcl-2 family proteins.

Objective: To longitudinally evaluate the outcome of secondary bone grafting (SBG) using computed tomograms (CTs) and conventional dental radiographs., Subjects: Nineteen alveolar clefts from 17 patients were used in this study., Method: A two-dimensional evaluation of SBG was performed using dental radiographs at 1 year after SBG by assigning scores of 1 to 4 (from very good to poor) based on postoperative marginal bone level on the alveolar side. On the basis of postoperative marginal bone levels on the nasal side, clefts were also assigned to groups with the bony bridge on or above (group I) or below (group II) a horizontal reference line. Three-dimensional evaluation of the SBG was performed on horizontal CT slices with the residual cortical bone (RCB) ratio before SBG (T0) as well as 1.5 (T1), 3 (T2), 6 (T3), and 12 months (T4) after SBG., Results: The RCB ratio at T4 in the group with scores 1 and 2 was significantly smaller than that of score 3. Furthermore, the mean RCB ratio at T4 in group I was significantly smaller than that in group II. Nineteen alveolar clefts were divided into two groups, A and B, based on a cluster analysis of the RCB ratios. Group A showed a continuous decrease in the RCB ratio from T0 to T2, but group B showed a significant decrease only in the period from T0 to T1., Conclusion: These results suggested that the RCB ratio might be a useful parameter for evaluation of the bony bridge after SBG.

A subcutaneous tumor localized on the sole of the foot of a 25-year-old Japanese man was diagnosed as a malignant melanoma of the soft parts. We performed a wide local excision, elective lymph node dissection, and chemotherapy. Eight years after treatment, the patient is alive without local recurrence or metastasis.

Various complications of Le Fort osteotomies have been reported. We describe a lethal complication of Le Fort III osteotomy we encountered in a 9-year-old boy with Crouzon syndrome. A standard Le Fort III osteotomy, including pterygomaxillary dysjunction with a curved osteotome and down-fracture manipulation, was performed uneventfully. When the intraoral buccal wound was closed after fixation of the external midface distraction devices, we discovered hemorrhage originating from the right posterior maxillary region. Although it was stopped with pressure on the osteotomized maxilla, the volume of intraoperative blood loss was nearly 2,000 ml. During the observation period in the intensive care unit, the patient suffered brain death, and he died 3 months later. A computed tomography scan obtained the day after surgery revealed vigorous subarachnoid and intraventricular hemorrhage and transverse fracture of the middle cranial fossa. This skull base fracture was believed to result from intraoperative maneuvers, including the pterygomaxillary dysjunction and down-fracture manipulation. We emphasize the risk of intracranial hemorrhage with Le Fort osteotomy and advise discussing this risk with patients and family members during preoperative consultations.

Background: Most cutaneous hemangiomas involute spontaneously. However, the lesion existing on the face poses a cosmetic problem during the waiting period for involution., Methods: Hemangioma showing late involution was treated by local injection of monoethanolamine oleate solution., Results: The lesion flattened and was excised serially., Conclusion: Sclerotherapy with monoethanolamine oleate can shorten the waiting period for involution and can decrease the risk of psychosocial trauma. Moreover, it provides good preparation for further surgery. Such sclerotherapy is effective in the treatment of hemangioma with late involution.

Objective: To examine the prognostic significance of the skeletal morphology around the nasal cavity and the alveolar cleft in secondary bone grafting (SBG)., Design and Setting: Fifty-one alveolar clefts in 41 patients (10 bilateral and 31 unilateral cleft lips and palates) registered in the Tokushima University Dental Hospital were examined in this study., Method: Evaluation of the bony bridge after SBG using dental radiographs at 1 year after surgery. The clefts were divided into two groups: group I (54.9%) in which the upper border of the bony bridge was preferably maintained on or above the horizontal reference line (RL) constructed at the level of the root apex of the upper central incisor adjacent to the cleft, and group II (45.1%) in which the bone level was lower than the RL. Presurgical cleft width was determined by the dental radiographs. The cleft/nasal cavity ratio; the value of the cleft width divided by the nasal cavity width on the cleft side, which was analyzed by frontal cephalograms before the SBG; and the cleft/apertura piriformis ratio, the value analyzed by computed tomography, were used., Results and Conclusion: The age, sex, and eruptive stage of the canine teeth at the time of the SBG showed no significant difference between groups. The presurgical cleft width also showed no significant difference between group I (6.6 +/- 3.1 mm) and group II (7.9 +/- 3.3 mm). The cleft/nasal cavity ratio showed a significant difference between groups I and II (0.42 +/- 0.14, 0.75 +/- 0.25; p < .05). Furthermore, the cleft/apertura piriformis ratio also showed a significant difference between groups I and II (0.32 +/- 0.12, 0.65 +/- 0.26; p < .05). These results suggested that measurements of the skeletal morphology around the nasal cavity and alveolar cleft might aid in predicting the stability of the bony bridge after SBG.

We treated midface hypoplasia in a 20-year-old woman with Crouzon syndrome using a rigid external distraction device. The patient showed severe exophthalmos and maxillary retrusion, although relatively good occlusion had been achieved by long-term orthodontic procedures. We considered that our patient's particular condition could not be resolved by the usual Le Fort III osteotomy/midface distraction procedure, so we devised a segmental approach. The midface, mobilized by Le Fort III osteotomy, was divided into two segments by Le Fort I osteotomy; each fragment was connected to the rigid external distraction device to be distracted separately. Distraction was begun after 1 day at 1 mm/day. The upper and lower segments were distracted over 17 and 12 days, respectively. The patient's occlusion was fully corrected, and her facial contour was significantly improved. After 3 weeks of consolidation, we removed the distraction device. The clinical course was without complication, and no relapse was observed on the cephalogram or computed tomography scan obtained 1 year after the procedure. Our modified technique was helpful in increasing the usefulness of the external distraction system and in refining the midface distraction procedure.

We used distraction osteogenesis to correct six deformed thumbs in four patients ranging in age from 4 to 7 years. Two of the patients had Apert syndrome (syndromic craniosynostosis with symmetrical syndactyly) and two had polydactyly. We used a small fixator with a ball joint and successfully corrected the angular deformity after lengthening the proximal phalanx by distraction. This single inclusive procedure was extremely useful. We found the optimal distraction regimen for the digital phalanx was a one day waiting period and lengthening at 1 mm/day. The mean healing indexes were 37.2 days/cm (range 24.2 to 41.5) in those with Apert syndrome and 64.3 days/cm in those with polydactyly (62.5 and 66.0). Our results suggest that osteogenesis at the distraction site may be quicker in patients with Apert syndrome than in those with polydactyly.

Background: Pyogenic granulomas have been treated by cryosurgery, curettage, electrodesiccation, chemical cauterization, and laser surgery. The therapeutic effects of these conservative treatments are limited, however., Objective: In this study, the efficacy of a sclerosing agent (monoethanolamine oleate) was evaluated for the treatment of pyogenic granuloma., Methods: Pyogenic granuloma was treated by local injection of the monoethanolamine oleate solution in nine patients who were from 1 to 57 years of age (median 18 years). The treatment effect was determined by physical examination., Results: All lesions were removed completely with no recurrence, and scars were inconspicuous in all cases. Complications occurred in only one patient who complained of pain due to an avoidable injection of excess solution., Conclusion: Sclerotherapy with monoethanolamine oleate is effective in the treatment of pyogenic granuloma and offers an alternative to conventional methods in cases for which conservative treatment is preferable.

We investigated the cutaneous blood supply in the gluteal and perineal regions of 35 donated cadavers to provide an anatomical basis for reliable vulvo-vaginal reconstruction using a skin flap such as the so-called gluteal fold flap. The cutaneous areas along the gluteal cleft and sulcus were likely to be supplied by 3 routes: 1) the internal pudendal artery (IPA), especially its first cutaneous branch; 2) perforators running through the gluteus maximus muscle and arising from the inferior gluteal artery (IGA); and 3) a non-perforator running around and inferior to the ischial tuberosity and originating from the IGA. Route 1 supplied the skin along the gluteal cleft, route 2 the gluteal fold (i.e., a bulky skin fold along the upper edge of the gluteal sulcus), and route 3, just along the gluteal sulcus. In those 3 routes, we noted the consistent morphology of the thick and long, first cutaneous branch of the IPA. The first arterial branch, 1.5 mm in diameter at its origin on average (ranging from 0.7-2.6 mm), usually originated from the IPA under the cover of or at the inferomedial or distal side of the sacrotuberous ligament (almost always less than 20 mm from the inferomedial margin of the ligament). The branch ran superomedially toward the coccyx or ran medially in the ischiorectal fat. It accompanied the vein and nerve at its distal (peripheral) course although the nerve often ran independently at its proxomal course near the ligament. Therefore, the first branch of the IPA seems to provide a reliable pedicle using the skin along the gluteal cleft whether the incision for approach is conducted along the gluteal sulcus or not. However, if the gluteus maximus muscle extended much inferomedially, the pedicle would be very short. In this case, preparation of the pedicle seems to be necessary along the arterial course under the cover of the muscle.

The case of a 1-year-old boy who underwent correction of a scaphocephalic deformity using distraction devices is described. Double pi-shaped osteotomies were cut. Bilateral temporal bone flaps were gradually expanded, and the frontal bone was pulled back simultaneously. The patient showed a good skull contour 2 years postoperatively. Although previous distractions were aimed only at expansion of the cranium, we used the distraction technique to successfully shorten the cranial vault in the anteroposterior direction. This case demonstrates that the osteotomized bone flaps can be moved in various directions. This feature of the distraction technique may contribute greatly toward meticulous reconstruction of the cranial vault.

We report a case of malignant melanoma arising on the hyperkeratotic sole of a patient with palmoplantar keratoderma (PPK). Hyperkeratotic lesions were also seen on the dorsa of both hands and feet and the extensor aspects of elbows and knees. The patient's PPK appeared to have been transmitted by an autosomal dominant gene. Histologically, the hyperkeratotic lesions showed acanthosis, marked hyperkeratosis without parakeratosis, and hypergranulosis. All the findings corresponded to those of Greither's disease with malignant melanoma.