Study on ammonia generation from digested sludge by subcritical water treatment.

Organic sludge has recently attracted attention as a renewable energy source. While the organic matter contained in sludge can be utilized as various renewable energy sources, its nitrogen component has limited use. In this study, subcritical water treatment was conducted to recover ammonia from digested sludge. While ammonia recovered via stripping is limited to soluble components, subcritical water treatment can convert solid components and dissolved organic nitrogen sludge into ammonia. Digested sludge was treated at several reaction temperatures, reaction pressures, treatment times, and oxygen ratios to determine the ammonia generation rate. Among the conditions tested in this study, an ammonium generation rate of 84.0% was obtained at 400 °C, 10 MPa, a treatment time of 5 min, and at an oxygen ratio of 1.2.

Effect of Te substitution on crystal structure and transport properties of AgBiSe2 thermoelectric material.

Silver bismuth diselenide (AgBiSe2) has attracted much attention as an efficient thermoelectric material, owing to its intrinsically low lattice thermal conductivity. While samples synthesized using a solid-state reaction showed n-type conductivity and their dimensionless figure of merit (ZT) reached ∼1 by electron doping, theoretical calculations predicted that a remarkably high thermoelectric performance can be achieved in p-type AgBiSe2. In this paper, we present the effect of Te substitution on the crystal structure and thermoelectric properties of AgBiSe2, expecting p-type conductivity due to the shallowing of the energy potential of the valence band. We found that all AgBiSe2-xTex (x = 0-0.8) prepared using a solid-state reaction exhibits n-type conductivity from 300 to 750 K. The room-temperature lattice thermal conductivity decreased to as low as 0.3 W m-1 K-1 by Te substitution, which was qualitatively described using the point defect scattering model for the solid solution. We show that ZT reaches ∼0.6 for x = 0.8 at a broad range of temperatures, from 550 to 750 K, due to the increased power factor, although the carrier concentration has not been optimized yet.

High-throughput powder diffraction measurement system consisting of multiple MYTHEN detectors at beamline BL02B2 of SPring-8.

In this study, we developed a user-friendly automatic powder diffraction measurement system for Debye-Scherrer geometry using a capillary sample at beamline BL02B2 of SPring-8. The measurement system consists of six one-dimensional solid-state (MYTHEN) detectors, a compact auto-sampler, wide-range temperature control systems, and a gas handling system. This system enables to do the automatic measurement of temperature dependence of the diffraction patterns for multiple samples. We introduced two measurement modes in the MYTHEN system and developed new attachments for the sample environment such as a gas handling system. The measurement modes and the attachments can offer in situ and/or time-resolved measurements in an extended temperature range between 25 K and 1473 K and various gas atmospheres and pressures. The results of the commissioning and performance measurements using reference materials (NIST CeO2 674b and Si 640c), V2O3 and Ti2O3, and a nanoporous coordination polymer are presented.

Usefulness of mandibular third molar coronectomy assessed through clinical evaluation over three years of follow-up.

The aim of this study was to investigate the 3-year morbidity of coronectomy of the lower third molar and to monitor the behaviour and migration pattern of the retained roots postoperatively. A total of 92 patients (111 teeth) who had undergone a coronectomy between October 2005 and July 2009 were investigated. Patients were followed up at 3 months and 1, 2, and 3 years for clinical evaluation and dental computed tomography imaging of the coronectomy sites. In total, 10 cases (9%) required tooth root extraction within the 3 years after coronectomy. In seven of them, the distal pocket of the lower second molars remained connected to the roots within the first year. Of the cases in whom a pocket did not remain at an early stage, none showed peri-apical lesions on transmission images of the retained roots in the apical area, which usually result from necrosis of the pulp. Root migration increased in the first 2 years after coronectomy but stabilized between the second and third years. In addition, a significant difference was noted in root migration between patients of different ages and sex. Retained roots after coronectomy in the lower third molars led to no complications in terms of infection or the development of pathologies within the first 3 years postoperatively.

Experimental verification of PbBi2Te4 as a 3D topological insulator.

The experimental evidence is presented of the topological insulator state in PbBi2Te4. A single surface Dirac cone is observed by angle-resolved photoemission spectroscopy with synchrotron radiation. Topological invariants Z2 are calculated from the ab initio band structure to be 1;(111). The observed two-dimensional isoenergy contours in the bulk energy gap are found to be the largest among the known three-dimensional topological insulators. This opens a pathway to achieving a sufficiently large spin current density in future spintronic devices.

Homozygous c.14576G>A variant of RNF213 predicts early-onset and severe form of moyamoya disease.

OBJECTIVE: RNF213 was recently reported as a susceptibility gene for moyamoya disease (MMD). Our aim was to clarify the correlation between the RNF213 genotype and MMD phenotype. METHODS: The entire coding region of the RNF213 gene was sequenced in 204 patients with MMD, and corresponding variants were checked in 62 pairs of parents, 13 mothers and 4 fathers of the patients, and 283 normal controls. Clinical information was collected. Genotype-phenotype correlations were statistically analyzed. RESULTS: The c.14576G>A variant was identified in 95.1% of patients with familial MMD, 79.2% of patients with sporadic MMD, and 1.8% of controls, thus confirming its association with MMD, with an odds ratio of 259 and p < 0.001 for either heterozygotes or homozygotes. Homozygous c.14576G>A was observed in 15 patients but not in the controls and unaffected parents. The incidence rate for homozygotes was calculated to be >78%. Homozygotes had a significantly earlier age at onset compared with heterozygotes or wild types (median age at onset 3, 7, and 8 years, respectively). Of homozygotes, 60% were diagnosed with MMD before age 4, and all had infarctions as the first symptom. Infarctions at initial presentation and involvement of posterior cerebral arteries, both known as poor prognostic factors for MMD, were of significantly higher frequency in homozygotes than in heterozygotes and wild types. Variants other than c.14576G>A were not associated with clinical phenotypes. CONCLUSIONS: The homozygous c.14576G>A variant in RNF213 could be a good DNA biomarker for predicting the severe type of MMD, for which early medical/surgical intervention is recommended, and may provide a better monitoring and prevention strategy.

Early diffusion MR imaging findings and short-term outcome in comatose patients with hypoglycemia.

BACKGROUND AND PURPOSE: The relationship between the MR imaging features and clinical outcomes in patients with hypoglycemic encephalopathy has always been evaluated retrospectively. The aim of this study was to prospectively evaluate whether MR imaging features of patients presenting with hypoglycemic coma are predictive of short-term (1-week) outcomes. MATERIALS AND METHODS: Subjects were 36 consecutive patients with hypoglycemia who were in a comatose state on arrival at our hospital from April 2006 to March 2010. MR imaging findings on arrival in relation to the patients' clinical course after glucose infusion were evaluated. RESULTS: Thirteen of the 36 patients showed no MR imaging abnormalities on arrival. DWI revealed focal lesions involving the internal capsule in 13 patients and lesions involving bilateral hemispheric white matter in 10 patients. After glucose administration, the patients without lesions and patients with focal internal capsule lesions recovered completely within 1 day. However, patients with diffuse white matter lesions did not recover even within 1 week despite glucose administration. There was no statistical difference in the initial blood glucose levels among patients with the various types of MR imaging findings. CONCLUSIONS: On early MR imaging, hypoglycemic brain injury may first appear in the internal capsule and then spread into the hemispheric white matter. The absence of a lesion or the presence of a focal internal capsule lesion may suggest a good outcome. However, diffuse hemispheric white matter lesions may indicate a poor 1-week outcome.

Positional nystagmus in patients with chronic dizziness.

BACKGROUND: In elderly people, chronic dizziness is endemic. However, chronic dizziness of unknown origin is difficult to assess. OBJECTIVE: To investigate whether mild unrecognised benign paroxysmal positional vertigo (BPPV) is a cause of isolated chronic dizziness in the elderly. PATIENTS AND METHODS: The prevalence of extremely weak, horizontal, direction changing apogeotropic positional nystagmus (HAPN) that had not been detected by conventional examination was evaluated in 200 patients with isolated chronic dizziness and in 155 age matched control subjects without dizziness. RESULTS: A high prevalence of weak HAPN was found in patients with isolated chronic dizziness (98/200 (49.0%)) compared with the prevalence in control subjects without dizziness (25/155 (16.1%); p<0.0001). Symptoms improved in some patients by daily positional exercise for BPPV. CONCLUSION: Because BPPV is the most common cause of dizziness in the elderly, and HAPN is a characteristic of horizontal canal BPPV, our findings suggest that mild persistent BPPV is a possible cause of chronic dizziness of otherwise unknown origin in the elderly.

Intra-bone marrow cotransplantation of donor mesenchymal stem cells in pig-to-NOD/SCID mouse bone marrow transplantation facilitates short-term xenogeneic hematopoietic engraftment.

We directly injected porcine donor mesenchymal stem cells (MSC) into murine bone marrow (BM) cavities to examine the effects of intra-BM cotransplantation of MSC in pig-to-NOD/SCID mouse bone marrow transplantation (BMT) on xenogeneic engraftment. Porcine MSC prepared by aspiration of iliac BM of miniature swine were identified as CD90+CD29+CD45-CD31- and shown to differentiate into osteoblastocytes and adipocytes. A few weeks after expansion, MSC (1 x 10(6) cells/mouse) were directly injected with BM cells (30 x 10(6) cells/mouse) obtained from vertebrae through a microsyringe into BM cavities of both tibiae of NOD/SCID mice after 3-Gy total body irradiation. Controls were injected with only BM cells. Porcine chimerisms of BM cells of tibiae (injection site) and of femurs (non-injection site) in recipient mice were evaluated with porcine and murine cell markers using FACS. The chimerism of porcine class I+ cells at the injection site in the MSC group and the controls were 3.45%, 1.43%, and 0.17%, and 2.27%, 0.81%, and 0.1% at 1, 3, and 6 weeks, respectively. The chimerism at the noninjection site in the MSC group and the controls were 0.21%, 1.34%, and 0.11%, and 0.06%, 0.42%, and 0.09% at 1, 3, and 6 weeks, respectively. The total chimerisms of injection site in the MSC group to 6 weeks were significantly higher than those in the control group (1.60% vs 0.99%; P < .05), whereas the chimerism of the noninjection site in MSC group was remarkably higher at 3 weeks. In conclusion, intra-BM cotransplantation of porcine donor MSC in pig-to-NOD/SCID mouse BMT improved short-term xenogeneic engraftment, presumably due to humoral factors.

High expression of GADD-45alpha and VEGF induced tumor recurrence via upregulation of IL-2 after photodynamic therapy using NPe6.

NPe6 is a novel second-generation photosensitizer used for photodynamic therapy (PDT). PDT using NPe6 and diode laser (664 nm) induces cell death, inflammatory reactions, immunological responses and damage to the microvasculature. In this study, we evaluated the influence of the immunological responses and of enhanced angiogenesis on the anti-tumor effect of NPe6-PDT using cytokine-overexpressing Lewis lung carcinoma (LLC), LLC-IL-2 cells both in vitro and in vivo. We showed by DNA microarray analysis in vitro that IL-2 and GADD-45alpha (growth arrest and DNA damage 45 alpha) mRNA expressions were induced by 3 h after NPe6-PDT applied at a dose killing 90% of the cells (LD90). IL-2-overexpressing cells (LLC/IL-2 cells) were resistant to the loss of clonogenicity as compared to the parental LLC cells in vitro. Furthermore, in female C57BL/6 mice, NPe6-PDT produced a cure rate of 66.7% in LLC tumors, whereas the cure rate was only 16.6% in LLC/IL-2 tumors, and overexpression of IL-2 caused failure of NPe6-PDT, with tumor recurrence, in vivo. These results suggest that IL-2 expression may play an unfavorable role in attenuation of the antitumor effect of NPe6-PDT. It has been reported that the expression of vascular endothelial growth factor (VEGF), in particular, may cause tumor recurrence after PDT and exert unfavorable effect in relation to attenuate the anti-tumor activity of PDT. Results of immunohistochemical analysis of LLC/IL-2 tumors have revealed that the expressions of GADD-45alpha and VEGF are induced in these tumors after PDT, and in particular, 12 h after PDT, the expression levels were much higher as compared with those in the LLC tumors. The results of our studies using in vitro and in vivo models suggest that the cell death caused by PDT was inhibited by induction of GADD-45alpha expression and that tumor recurrence was promoted by the enhancement of VEGF expression mediated by IL-2 upregulation. Therefore, it is speculated that the use of an IL-2 inhibitor may improve the efficacy of NPe6-PDT.

Safety assessment of ellagic acid, a food additive, in a subchronic toxicity study using F344 rats.

Ellagic acid is a phenolic acid compound, used as a food additive for its antioxidative properties. Because of its chemical characteristics, use is also to be expected in cosmetics. The present 90-day subchronic toxicity study was performed in F344 rats at dose levels of 0, 1.25, 2.5 and 5% in powdered basal diet, with actual doses of 9.4, 19.1, 39.1 g/kg b.w., respectively, in males, and 10.1, 20.1, 42.3 g/kg b.w. in females. No mortality or treatment-related clinical signs were observed throughout the experimental period. Body weight gain was significantly reduced from weeks 3 (5% group), 6 (2.5% group) and 7 (1.25% group) to the end of the experiment (except week 8 in the lowest group) in the treated females, the final body weights being decreased in the 5% (92.5%), 2.5% (94.2%) and 1.25% (94.8%) treated groups as compared to the control. Changes in MCV and serum AST, ALP, Ca, Cl and P were sporadically observed, but these were not considered to be treatment-related alterations. There were no obvious histopathological changes in any of the groups. The no-observed-effect level (NOEL) was estimated to be 5% (3011 mg/kg b.w./day) for males and the no-observed-adverse-effect level (NOAEL) and NOEL in females were estimated to be 5% (3254 mg/kg b.w./day) and <1.25% (778 mg/kg b.w./day), respectively.

A possible relationship between the anti-cancer potency of photodynamic therapy using the novel photosensitizer ATX-s10-Na(II) and expression of the vascular endothelial growth factor in vivo.

ATX-s10-Na(II) is a novel second-generation photo-sensitizer for photodynamic therapy (PDT). PDT using ATX-s10 and diode laser (670 nm) induces an apoptotic response, inflammatory reaction, immune reaction and damage to the microvasculature. In particular, the vascular shut-down effect plays an important role in the anti-tumor activity of ATX-s10-PDT. It has been reported that PDT induces hypoxia and expression of the vascular endothelial growth factor (VEGF) via the hypoxia-inducible factor 1 (HIF1)-alpha pathway. We hypothesized that the expression of VEGF may cause tumor recurrence after PDT and exert unfavorable effect against the anti-tumor activity of ATX-s10-PDT. In this study, we showed by DNA microarray analysis in vitro that VEGF mRNA expression was induced 3 h after laser irradiation in ATX-s10-PDT. We compared the anti-tumor activity of ATX-s10-PDT against lung cancer cell lines SBC-3 and SBC-3/VEGF, the latter overexpressing VEGF; there was no significant difference in the sensitivity to the PDT between the two cell lines as assessed by clonogenic assay. Furthermore, no statistically significant difference in the anti-tumor effect of PDT, as measured by tumor cures, was found between SBC-3 and SBC-3/VEGF tumors in female Balb/c-nu/nu nude mice in vivo. In conclusion, ATX-s10-PDT may prevent tumor recurrence despite induction of VEGF and promotion of tumor angiogenesis, which are known to enhance tumor proliferation and survival.

Transgenic animal production and animal biotechnology.

Considerable progress has been made in methods for production of transgenic livestock; beginning with pronuclear microinjection over 20 years ago. New methods, including the use of viral vectors, sperm-mediated gene transfer and somatic cell cloning, have overcome many of the limitations of pronuclear microinjection. It is now possible to not only readily make simple insertional genetic modifications, but also to accomplish, more complex, homozygous gene targeting and artificial chromosome transfer in livestock.

L-3,4-dihydroxyphenylalanine-induced c-Fos expression in the CNS under inhibition of central aromatic L-amino acid decarboxylase.

L-3,4-dihydroxyphenylalanine (DOPA) is a neurotransmitter candidate. To map the DOPAergic system functionally, DOPA-induced c-Fos expression was detected under inhibition of central aromatic L-amino acid decarboxylase (AADC). In rats treated with a central AADC inhibitor, DOPA significantly increased the number of c-Fos-positive nuclei in the paraventricular nuclei (PVN) and the nucleus tractus solitarii (NTS), and showed a tendency to increase in the supraoptic nuclei (SON), but not in the striatum. On the other hand, DOPA with a peripheral AADC inhibitor elevated the level of c-Fos-positive nuclei in the four regions, suggesting that DOPA itself induces c-Fos expression in the SON, PVN and NTS. In rats treated with 6-hydroxydopamine (6-OHDA) to lesion the nigrostriatal dopamine (DA) pathway, DOPA significantly induced c-Fos expression in the four regions under the inhibition of peripheral AADC. However, under the inhibition of central AADC, DOPA did not significantly increase the number of c-Fos-positive nuclei in the four regions, suggesting that DOPA at least in part induces c-Fos expression through its conversion to DA. It was likely that the 6-OHDA lesion enhanced the response to DA, but attenuated that to DOPA itself. In conclusion, we proposed that the SON, PVN and NTS include target sites for DOPA itself.

A subchronic toxicity study of dunaliella carotene in F344 rats.

Dunaliella carotene, extracted from dunaliella alga (Dunaliella bardawil or Dunaliella salina), for use as a food-coloring agent, has beta-carotene as its mainly constituent. As there have been no reports of toxicological evaluation, a 90-day subchronic toxicity study was here performed in F344 rats at dose levels of 0 (control), 0.63%, 1.25%, 2.5% and 5% in powdered basal diet. The average daily intakes of dunaliella carotene were 352, 696, 1420 and 2750 mg/kg/day, respectively, for males, and 370, 748, 1444 and 2879 mg/kg/day for females. No mortality or treatment-related clinical signs were observed throughout the experimental period in any of the groups. Body weight gain was slightly but significantly (p < 0.05) reduced from week 5 to the end of the experiment in 2.5% and 5% males. Increased PLT were observed in 1.25% and 5% males, and 2.5% and 5% females. Significant elevations or tendencies for increase in serum T. Cho and Ca were observed in all treated males and females, with clear dose-dependence in males. Organ weight measurement and histopathological observation revealed no toxicological changes. Based on growth suppression, no-observed-adverse-effect-levels (NOAELs) were estimated to be 1.25% (696 mg/kg/day) for males and 5% (2879 mg/kg/day) for females. As increases in serum Ca were observed in the lowest group in both sexes, a no-observed-effect level (NOEL) could not be determined in this study.

Lack of carcinogenicity of D-xylose given in the diet to F344 rats for two years.

The two year carcinogenicity of D-xylose was examined in groups of 50 male and 50 female F344 rats at dietary doses of 0% (control), 2.5% and 5%. The doses were selected on the basis of results from a 13-week subchronic toxicity study. Growth suppression and soft feces were observed in male and female rats of the 5% group. However, no significant differences from the controls were noted with regard to clinical signs, mortality and hematological findings. Decrease in absolute weight and increase in relative weight of the brain in males, and decrease of absolute kidney weight in females were observed in the 5% group, but there were no remarkable histopathological changes. A variety of tumors developed in all groups, including the controls, but all were histologically similar to those known to occur spontaneously in F344 rats, and no statistically significant increase in the incidence of any type of neoplastic lesion was found for either sex in the treated groups. Thus, it was concluded that, under the present experimental conditions, D-xylose is not carcinogenic to F344 rats.

Lack of subchronic toxicity of an aqueous extract of Agaricus blazei Murrill in F344 rats.

Agaricus blazei Murrill, an edible mushroom, is widely used as a functional food due to its possible medicinal effects. Aqueous extracts are also used as food additive to provide an agreeable bitter taste. As a part of its safety assessment, the present 90-day subchronic toxicity study was performed in F344 rats. To establish a no-observed-adverse-effect level (NOAEL), rats were fed powder diet containing A. blazei Murrill aqueous extract at dose levels of 0 (basal diet), 0.63, 1.25, 2.5 and 5% (maximum) for 90 days. During the experiment, there were no remarkable changes in general appearance and no deaths occurred in any experimental group. Although serum blood urea nitrogen was slightly but significantly increased in males of the 2.5 and 5% groups, no related histopathological changes were observed in the kidney, and serum creatinine levels were rather reduced, suggesting the increase of blood urea nitrogen to be of little toxicological significance. Hematology, organ weight measurement and histopathological observation revealed no test compound-related toxicological changes. In conclusion, A. blazei Murrill extract even at 5% in the diet (2654 mg/kgb.w./day for male rats and 2965 mg/kgb.w./day for female rats) did not cause remarkable adverse effects in F344 rats. Thus, the NOAEL was concluded to be 5% in the diet.

Human artificial chromosome (HAC) vector provides long-term therapeutic transgene expression in normal human primary fibroblasts.

Human artificial chromosomes (HACs) segregating freely from host chromosomes are potentially useful to ensure both safety and duration of gene expression in therapeutic gene delivery. However, low transfer efficiency of intact HACs to the cells has hampered the studies using normal human primary cells, the major targets for ex vivo gene therapy. To elucidate the potential of HACs to be vectors for gene therapy, we studied the introduction of the HAC vector, which is reduced in size and devoid of most expressed genes, into normal primary human fibroblasts (hPFs) with microcell-mediated chromosome transfer (MMCT). We demonstrated the generation of cytogenetically normal hPFs harboring the structurally defined and extra HAC vector. This introduced HAC vector was retained stably in hPFs without translocation of the HAC on host chromosomes. We also achieved the long-term production of human erythropoietin for at least 12 weeks in them. These results revealed the ability of HACs as novel options to circumvent issues of conventional vectors for gene therapy.