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Rat hepatitis E virus (ratHEV) genome has four open reading frames (ORFs: ORF1, ORF2, ORF3 and ORF4). The functions of ORF3 and ORF4 are unknown. An infectious cDNA clone (pUC-ratELOMB-131L_wt, wt) and its derivatives including ORF3-defective (ΔORF3) and ORF4-defective (ΔORF4) mutants, were constructed and their full-length RNA transcripts transfected into PLC/PRF/5 cells. ΔORF3 replicated as efficiently as wt in cells. However, ≤1/1000 of the number of progenies were detectable in the culture supernatant of ΔORF3-infected cells compared with wt-infected cells. ORF4 protein was not detectable in ratHEV-infected cells or in the liver tissues of ratHEV-infected rats. No marked differences were noted between wt and ΔORF4 regarding the viral replication and protein expression. ORF3 mutants with proline-to-leucine mutations at amino acids (aa) 93, 96 and/or 98 in ORF3 were constructed and transfected into PLC/PRF/5 cells. Wt and an ORF3 mutant with leucine at aa 98 (ORF3-L98) replicated efficiently (density 1.15–1.16 g/cm 3 ), while ORF3-L93 + L96 exhibited a decreased viral release and banded at 1.26–1.27 g/cm 3 , similar to ΔORF3. In conclusion, the ORF3 protein, especially its proline residues at aa 93 and 96, is essential for the release of membrane-associated ratHEV particles, and ORF4 is unnecessary for the replication of ratHEV. [ABSTRACT FROM AUTHOR]

Hepatitis E virus (HEV) causes acute or chronic hepatitis in humans and can be transmitted via the fecal-oral route. Pigs are one of the main reservoirs for this infection. Sixty pigs, 4–5 months of age, on a swine herd in Japan had detectable anti-HEV IgG antibodies, and five (8.3%) of them had ongoing infection of genotype 3 HEV. Five HEV strains obtained from the viremic pigs shared 98.8–100% nucleotide identity, and one representative strain (swHE1606845), whose entire genomic sequence was determined in this study, differed by 14.1–19.6% from the reported HEV strains of subtypes 3a–3k and by 14.7–19.1% from other genotype 3 HEV strains whose subtypes have not yet been assigned. swHE1606845 showed a higher nucleotide p-distance value of ≥0.143 with the genotype 3 HEV strains of subtypes 3a–3k and ≥0.152 with other genotype 3 strains of unassigned subtypes. A SimPlot analysis revealed a lack of recombination events. These results indicate that swHE1606845 is a candidate member of a novel subtype of genotype 3. Further efforts to identify the swHE1606845-like novel strain are warranted to clarify the origin of this strain and to determine the complete nucleotide sequences of two additional swHE1606845-like strains for assigning a new subtype. [ABSTRACT FROM AUTHOR]

We demonstrate a general process for fabricating graphene nanoelectronic devices that have next several features: free-standing, micrometer-sized monolayer graphene with high quality, arbitrarily-shaped metallic electrodes or sensors. In contrast to the normal routes, a gas etching process is used to create a deep trench in silicon for suspending the whole graphene device in a much larger area. User-designed electrodes or sensors are fabricated on the suspended graphene at the same time for realizing multiple functions. In this work, a suspended gold nanofilm sensor is designed to measure the intrinsic electrical and thermal properties of graphene on site. The sensor serves as both electrode and precise resistance thermometer at the same time. By simply changing the metallic electrode shape and electrical circuit, the free-standing graphene can be made into different devices, such as single-molecule detector or nano-resonator. In order to test the robustness of graphene device, a high electrical current is applied to heat the graphene in vacuum until it breaks. The breakdown current density is measured to be 1.86 mA/μm. More importantly, this method is not only limited to graphene, but also can be applied to any other two-dimensional materials. [ABSTRACT FROM AUTHOR]

Pancreatic cancer is recalcitrant to treatment as it is highly metastatic and rapidly progressive. While observing the behavior of human pancreatic BxPC-3 cells using an optical assay device called TAXIScan, we found that several synthetic pyrazole and pyrimidine derivatives inhibited cell migration. One such compound, 14–100 , inhibited metastasis of fluorescence-labeled BxPC-3 cells, which were transplanted into the pancreas of nude mice as a subcutaneously grown cancer fragment. Surprisingly, despite its low cytotoxicity, the compound also showed an inhibitory effect on cancer cell proliferation in vivo, suggesting that the compound alters cancer cell characteristics needed to grow in situ. Single-cell RNA-sequencing revealed changes in gene expression associated with metastasis, angiogenesis, inflammation, and epithelial-mesenchymal transition. These data suggest that the compound 14–100 could be a good drug candidate against pancreatic cancer. [Display omitted] • Synthesized pyrazole compound 14–100 inhibits pancreatic cancer cell growth in vivo. • Angiogenic ability and HSP70 expression is altered by 14–100. • Expression of CXCL8 was downregulated in the 14–100 treated cancer cells. • 14–100 has lower toxicity compared to the approved drug for pancreatic cancer. [ABSTRACT FROM AUTHOR]

Concise syntheses of 9-methoxyellipticine, 3,4-dihydroellipticine (µ-alkaloid D), 1,2,3,4-tetrahydroellipticine, 2-methyl-1,2,3,4-tetrahydroellipticine, olivacine, 3,4-dihydroolivacine, (±)-guatambuine, and (±)-janetine were developed starting from hexatriene intermediates readily obtained by Pd-catalyzed tandem cyclization/cross-coupling reaction of indolylborates. The route enables the facile construction of pyrido[4,3- b]carbazoles by Cu-catalyzed 6π-electrocyclization and subsequent transformation of the pyridocarbazole intermediates into pyrido[4,3- b]carbazole alkaloids. [ABSTRACT FROM AUTHOR]

As a solid-state membrane with only one-atom thickness, nano-porous graphene has attracted intense attention in many critical applications. Here, the key challenge is to suspend a single-layer graphene (SLG) and drill nanopores with precise dimensions. Here, we report a simple and reliable route for making suspended fluorinated SLG with size-tunable nanopores. Our method consists of two steps: 1. a free-standing SLG ribbon was created between two gold pads after deep dry etching of silicon substrate by xenon difluoride. The SLG was fluorinated by 5–13%. Superior to the normal wet etching method, the dry etching process is much simpler and results in less hole-defect and edge deformation. A large area fluorinated SLG can be suspended due to the sufficient etch depth. 2. a focused ion beam was introduced to drill nanopores in graphene with an initial diameter around 20 nm. Followed by an electron beam induced carbon deposition, the diameter of nanopore was gradually decreased to sub-10 nm. By changing the deposition time, the size of nanopore can be precisely controlled. High-cost transmission electron microscope is no longer needed. Our method provides a simple and effective way for preparing free-standing fluorinated SLG ribbon suitable for single-molecule detection. [ABSTRACT FROM AUTHOR]

Thermoresponsive properties of poly(7-methacryloyloxycoumarin) ( P1a ) and its derivatives as homopolymers containing a photoreactive coumarin unit were systematically investigated. P1a showed a lower critical solution temperature (LCST)-type phase separation in dichloromethane, chloroform, and 1,1,2-trichloroethane. The temperature producing a 50% transmittance ( T c ) of 0.1 wt% P1a in distilled chloroform was 26 °C, while the T c value decreased to 21 °C in deuterated chloroform i.e., the deuterium isotope effect. The T c values in chloroform were adjustable from 28 to 57 °C by the introduction of the ethyleneoxy spacer and the substitution of α-methyl group by hydrogen. While the polymers containing 4-substituted coumarins were soluble in chloroform and dichloromethane from 0 °C to their boiling points. In the distilled chloroform solution, the coumarin units in P1a underwent a [2 + 2] cycloaddition and the transmittance at 500 nm decreased from 98% to 29% at 25 °C by a 180-s photoirradiation (326 mJ/cm 2 , emission band at 365 nm). [ABSTRACT FROM AUTHOR]

To accurately characterize the shape of interfacial nanobubbles using atomic force microscopy (AFM), we investigated the effect of wettability of the AFM tip while operating in the peak force tapping (PFT) mode. The AFM tips were made hydrophobic and hydrophilic by Teflon AF coating and oxygen plasma treatment, respectively. It was found that the measured base radius of nanobubbles differed between AFM height images and adhesion images, and that this difference depended on the tip wettability. The force curves obtained during the measurements were also different depending on the wettability, especially in the range of the tip/nanobubble interaction and in the magnitude of the maximum attractive force in the retraction period. The difference suggests that hydrophobic tips penetrate the gas/liquid interface of the nanobubbles, with the three phase contact line being pinned on the tip surface; hydrophilic tips on the other hand do not penetrate the interface. We then quantitatively estimated the pinning position and recalculated the true profiles of the nanobubbles by comparing the height images and adhesion images. As the AFM tip was made more hydrophilic, the penetration depth decreased and eventually approached zero. This result suggests that the PFT measurement using a hydrophilic tip is vital for the acquisition of reliable nanobubble profiles. [ABSTRACT FROM AUTHOR]

The O 4 -benzo[ c ]phenanthridine alkaloids exhibit potent antiproliferative activity against cancer cells, which is derived from their ability to inhibit of topoisomerase I and II. It has been reported that in the alkaloids a cationic quaternary ammonium atom, which results in resonance effects between ring A and B, is necessary for increased antiproliferative activity. These findings indicate the role of their substituents at ring A on inhibition of tumor cell proliferation. In the present study, we systematically assessed the cytotoxic activities of naturally occurring alkaloids and their derivatives containing various ring A substituents against two tumor cell lines, HCT-116 colon tumor cells and HL-60 promyelocytic leukemia cells. Among the cationic iminium alkaloids, which displayed more potent activity than the corresponding neutral derivatives, and the 7,8-oxygenated benzo[ c ]phenanthridine alkaloids, chelerythrine and NK109, exhibited stronger antiproliferative activity than the 8,9- and 9,10-oxygenated alkaloids. The activity of cationic iminium alkaloids could be correlated with the bond lengths of their ring A substituents and the electrostatic potentials of their ammonium molecules by DFT calculation. [ABSTRACT FROM AUTHOR]

The relationship between the chemical structure of PMMA-type polymers and the removal rate by hydrogen radicals was examined. Hydrogen radicals are known to have superior reduction ability and can be generated by decomposing H 2 on hot-wire surfaces. PMMA-type polymers, which are widely used as base polymers for ArF resist, have many variations with the difference in the substituents. The removal rate of the main chain scission type polymers was faster than that of the main chain cross-linking type polymers. In the main chain cross-linking type polymers, it is considered that reduction decomposition of polymer was inhibited apparently according to occurrence to simultaneously reduce decomposition by hydrogen radicals and main chain cross-link. The removal rate was slow in the case of the main chain scission type polymers with a benzene ring. We guessed that the polymer with a benzene ring was a stable structure by resonance stabilization of a benzene ring. We found that the removal rate was slow according to increasing bulkiness of α-side chain in the main chain scission type polymers. It is considered that etching resistance increases with increasing bulkiness of α-side chain of polymer. [ABSTRACT FROM AUTHOR]

Objective Serum uric acid (SUA) levels in humans are mainly regulated by urate transporters. Recent genome-wide association studies suggested that common variants of the human sodium-dependent phosphate cotransporter type 1 gene ( NPT1/SLC17A1) influence SUA. NPT1 has been reported to mediate urate transport, but its physiologic role in regulating SUA in humans remains unclear. Furthermore, the findings of replication studies of the relationship between NPT1 variants and gout have been inconsistent. The aims of this study were to investigate the effect of NPT1 on gout and to determine its physiologic role. Methods Five hundred forty-five male Japanese patients with gout and 1,115 male Japanese control subjects were genotyped for rs1165196 (I269T), a common missense variant in NPT1. Analyses of the association between rs1165196 and gout were then conducted, focusing especially on renal underexcretion (RUE) gout. Immunohistochemical analysis and functional analysis using Xenopus oocytes were also performed. Results Single-nucleotide polymorphism rs1165196 significantly decreased the risk of RUE gout (odds ratio 0.73, P = 0.031) but did not confer a risk for all gout ( P = 0.123). The immunohistochemical analysis revealed that human NPT1 is localized to the apical membrane of the renal proximal tubule. The functional analysis using Xenopus oocyte expression systems showed that rs1165196 increases NPT1-mediated urate export. Conclusion This study showed that NPT1 is a urate exporter located in the renal proximal tubule in humans, and that its common gain-of-function variant, rs1165196, causes RUE gout, a major subtype of gout. These findings enable us to deepen our understanding of the physiologic role of NPT1 as a renal urate exporter as well as its pathophysiologic role in gout. [ABSTRACT FROM AUTHOR]

Abstract: This paper discusses the removal of chemically amplified negative-tone i-line resist SU-8 using hydrogen radicals generated by the catalytic decomposition of H2 molecules in H2/N2 mixed gas (H2:N2 =10:90vol.%) using a tungsten hot-wire catalyzer. SU-8 resists with exposure doses from 7 to 280mJ/cm2 were removed by hydrogen radicals, although the SU-8 removal rate was independent of the exposure dose. The SU-8 removal rate increased with both substrate and catalyzer temperature, in addition to a decrease in the distance between the catalyzer and substrate. A high removal rate for cross-linked SU-8 with an exposure dose of 14mJ/cm2 of approximately 4μm/min was achieved with a catalyzer to substrate distance of 20mm, and catalyzer and initial substrate temperatures of 2400 and 165°C, respectively. [Copyright &y& Elsevier]

Highlights: [•] A novel surface temperature profiler using a single carbon nanotube is developed. [•] The quantitative temperature measurement is validated. [•] Sensor sensitivity is analyzed and the sensor configuration is optimized. [•] Heating technique at the nanoscale contact point is developed. [Copyright &y& Elsevier]

Boron neutron capture therapy (BNCT) is a cancer treatment with clinically demonstrated efficacy using boronophenylalanine (BPA) and sodium mercaptododecaborate (BSH). However, tumor tissue selectivity of BSH and retention of BPA in tumor cells is a constant problem. To ensure boron accumulation and retention in tumor tissues, we designed a novel polyethylene glycol (PEG)-based boron-containing lipid (PBL) and examined the potency of delivery of boron using novel PBL-containing liposomes, facilitated by the enhanced permeability and retention (EPR) effect. PBL was synthesized by the reaction of distearoylphosphoethanolamine and BSH linked by PEG with Michael addition while liposomes modified using PBL were prepared from the mixed lipid at a constant molar ratio. In this manner, novel boron liposomes featuring BSH in the liposomal surfaces, instead of being encapsulated in the inner aqueous phase or incorporated in the lipid bilayer membrane, were prepared. These PBL liposomes also carry additional payload capacity for more boron compounds (or anticancer agents) in their inner aqueous phase. The findings demonstrated that PBL liposomes are promising candidates to effect suitable boron accumulation for BNCT. [ABSTRACT FROM AUTHOR]

( S)-(-)-Dichotomine A and its enantiomer were synthesized from the key intermediate, methyl 1-(1-hydroxyethyl)-β-carboline-3-carboxylate, by enantioselective esterification with Lipase QLM. The first total synthesis of (+)-dichotomide II and its enantiomer were also achieved from ( S)-(-)-dichotomine A methyl ester and its enantiomer. The absolute configuration of the stereogenic center of the reported (+)-dichotomide II was determined to be R. [ABSTRACT FROM AUTHOR]

The sporulation inhibitory gene spi in the pock-forming conjugative plasmid pSA1.1 of Streptomyces azureus was introduced into cells via a high or low copy number vector to examine the effect of gene dosage on the growth of Streptomyces lividans TK24 as a host. In transformants carrying a high spi copy number, nutrient mycelial growth was inhibited, as was morphological differentiation from substrate mycelium to aerial mycelium on solid media. The degree of inhibition depended on the spi gene dosage, but the presence of pSA1.1 imp genes, which encode negative repressor proteins for spi, relieved the inhibition. Confocal images of Spi tagged with enhanced green fluorescent protein in cells on solid media revealed that spi expression was initiated at the time of elongation of substrate mycelium, that its expression increased dramatically at septation in aerial hyphae, and that the expression was maximal during prespore formation. Expression of spi covered the whole of the hyphae, and the level of expression at the tip of the hyphae during prespore formation was about sixfold greater than during substrate mycelial growth and threefold greater than during aerial mycelial growth. Thus, localized expression of spi at particular times may inhibit sporulation until triggering imp expression to repress its inhibitory effects. [ABSTRACT FROM AUTHOR]

Abstract: Anaerobic ammonium oxidation (anammox) and denitrification are two distinct microbial reactions relevant to the global nitrogen cycle. The proposed initial step of the anammox reactions, reduction of nitrite to nitric oxide, has been postulated to be identical to that in denitrification catalyzed by the dissimilatory nitrite reductase of the cytochrome cd 1-type. Here, we characterized the copper-containing nitrite reductase homolog encoded by nirK detected in the genome of an anammox bacterium strain KSU-1. We hypothesize that this NirK-type nitrite reductase, rather than a nitrite reductase of the cytochrome cd 1-type (NirS), is likely to catalyze nitrite reduction in anammox organism KSU-1. [Copyright &y& Elsevier]

Mutations in the apically located Na+-dependent phosphate (NaPi) cotransporter, SLC34A3 (NaPi- IIc), are a cause of hereditary hypophosphatemic rickets with hypercalciuria (HHRH). We have characterized the impact of several HHRH mutations on the processing and stability of human NaPi-IIc. Mutations S138F, G196R, R468W, R564C, and c.228delC in human NaPi-IIc significantly decreased the levels of NaPi cotransport activities in Xenopus oocytes. In S138F and R564C mutant proteins, this reduction is a result of a decrease in the Vmax for Pi, but not the Km. G196R, R468W, and c.228delC mutants were not localized to oocyte membranes. In opossum kidney (OK) cells, cell surface labeling, microscopic confocal imaging, and pulse-chase experiments showed that G196R and R468W mutations resulted in an absence of cell surface expression owing to endoplasmic reticulum (ER) retention. G196R and R468W mutants could be partially stabilized by low temperature. In blue native-polyacrylamide gel electrophoresis analysis, G196R and R468W mutants were either denatured or present in an aggregation complex. In contrast, S138F and R564C mutants were trafficked to the cell surface, but more rapidly degraded than WT protein. The c.228delC mutant did not affect endogenous NaPi uptake in OK cells. Thus, G196R and R468W mutations cause ER retention, while S138F and R564C mutations stimulate degradation of human NaPi-IIc in renal epithelial cells. Together, these data suggest that the NaPi-IIc mutants in HHRH show defective processing and stability. [ABSTRACT FROM AUTHOR]

Electron-beam-induced deposition (EBID) is a simple and versatile technique of processing materials for three-dimensional nanoscale structures. A variety of precursor molecules can be used to build a localized solid deposition by the exposure of a substrate to an electron beam. This paper reports nano carbon deposition using solid n-tetracosane as a precursor, because this paraffin-based EBID can be introduced into existing scanning electron microscope (SEM) systems without difficulty. The paraffin is prepared on an aluminum film and operated by a manipulator in the SEM. The effects of the accelerating voltage, beam current, magnification, distance from the paraffin to the exposure point, the amount of paraffin, and the working distance are measured and discussed. It is found that the electron-beam-induced etching and the beam diameter are sometimes the dominant factors influencing the deposition rate, and that the thickness of the paraffin also affects the deposition distribution. Electron-beam bending, which is a critical factor causing degradation of nanofabrication, is treated carefully in order to understand the declination of carbon pillars. The obtained carbon pillar configurations suggest that electrically charged paraffin produced during preliminary irradiation of the electron beam causes a Coulomb force and results in the movement effect of deposition. © 2011 Wiley Periodicals, Inc. Electron Comm Jpn, 94(5): 39-46, 2011; Published online in Wiley Online Library (). DOI 10.1002/ecj.10192 [ABSTRACT FROM AUTHOR]

A dimeric cytochrome c with an apparent molecular mass of 25 kDa was isolated from an anammox bacterium, strain KSU-1, in a relatively large quantity. This protein was named the NaxLS complex. The spectrum of the oxidized form exhibited a peculiar Soret peak at 419 nm. The reduction of NaxLS was not complete even with the addition of excess dithionite, but was complete with titanium (III) citrate, indicating that the NaxLS complex has a very low redox potential. The genes encoding the two subunits, naxL and naxS, are adjacent on the genome. The deduced amino-acid sequences of the genes showed high identities with those of two genes encoding 'unknown proteins' in the genome of Candidatus Kuenenia stuttgartiensis, but had lower identities with other c-type heme proteins. The electron paramagnetic resonance spectra of NaxLS exhibited low-spin signals of two heme species in the range between g=2.6 and g=1.8, which strongly suggested an unusual His/Cys coordination. This unique coordination might account for the low redox potential of the hemes in NaxLS. NaxLS might participate in the transfer of low redox potential electrons in the intracellular anammoxosome compartment or the cytoplasm. [ABSTRACT FROM AUTHOR]

The type IIc Na+-dependent phosphate cotransporter (NaPi-IIc) is specifically targeted to, and expressed on, the apical membrane of renal proximal tubular cells and mediates phosphate transport. In the present study, we investigated the signals that determine apical expression of NaPi-IIc with a focus on the role of the N- and the C-terminal tails of mouse NaPi-IIc in renal epithelial cells [opossum kidney (OK) and Madin-Darby canine kidney cells]. Wild-type NaPi-IIc, the cotransporter NaPi-IIa, as well as several IIa-IIc chimeras and deletion mutants, were fused to enhanced green fluorescent protein (EGFP), and their cellular localization was analyzed in polarized renal epithelial cells by confocal microscopy and by cell-surface biotinylation. Fluorescent EGFP-fused NaPi-IIc transporter proteins are correctly expressed in the apical membrane of OK cells. The apical expression of N-terminal deletion mutants (deletion of N-terminal 25, 50, or 69 amino acids) was not affected by truncation. In contrast, C-terminal deletion mutants (deletion of C-terminal 45, 50, or 62 amino acids) did not have correct apical expression. A more detailed mutational analysis indicated that a domain (amino acids WLHSL) in the cytoplasmic C terminus is required for apical expression of NaPi-IIc in renal epithelial cells. We conclude that targeting of NaPi-IIc co the apical cell surface is regulated by a unique amino acid motif in the cytoplasmic C-terminal domain. [ABSTRACT FROM AUTHOR]

Abstract: In this study, a swim-bed reactor was used for partial nitrification treatment of magnesium ammonium phosphate precipitation process treated effluent of methane fermentation digester liquor. Even without temperature, DO, and pH control, the maximum nitrite production rate of 2.66kg NO2–N/m3/d was achieved under a nitrogen loading rate of 5.45kgN/m3/d at 18°C and a nitrite production rate of 0.22kg NO2–N/m3/d was still attained at a nitrogen loading rate of 0.48kgN/m3/d even the operational temperature decreased to 8°C. The average effluent NO2–N/(NO2 +NO3–N) ratio reached 95.4% during the entire experimental period. The mean particle size of suspended sludge increased from 260 to 450μm and the microscopic observation confirmed the formation of dense-flocs. The results indicated that there were two regimes for ammonium oxidation, one at 20–30°C and the other at 10–20°C. Scanning electron microscopic observation and FISH analysis demonstrated that ammonium-oxidizing bacteria were the dominant microorganisms and were considered to be responsible for the satisfactory partial nitrification performance. [Copyright &y& Elsevier]

Abstract: Underground brine waste containing high concentrations of ammonium and with a salinity of 3% is usually generated during the production of methane gas and iodine in the gas field of Chiba Prefecture, Japan. In this study, one swim-bed reactor, packed with a novel acrylic fiber biomass carrier (Biofringe), was applied to the partial nitritation treatment of this kind of underground brine waste. A stable nitrite production rate of 1.6 kg NO2-N m−3 d−1 was obtained under a nitrogen loading rate of 3.0 kg-N m−3 d−1, at a pH of 7.5 and a temperature of 25 °C. Nitrate production was negligible and the effluent NO2-N/NO x -N ratio was above 98% due to the successful inhibition of nitrite-oxidizing bacterial activity. Free ammonia was considered to be the main factor for inhibiting the activity of nitrite-oxidizing bacteria. A microbial community shift was demonstrated by 16S rRNA analysis, and it was shown that the ammonium-oxidizing bacteria became the predominant species after successful nitrite accumulation was observed. [Copyright &y& Elsevier]

The anammox process, as an alternative to conventional nitrogen removal technologies, has abstracted much attention in recent years. In this study, one column-type reactor using a novel support material--net type acrylic fiber (Biofix)--was used for anammox treatment. The Biofix reactor was operated at 25°C (peak summer temperature, 31.5°C). Over 330 days of operation for synthetic wastewater treatment, the nitrogen loading rates of the reactor were increased to 3.6 kg-N/m3/d and T-N removal efficiencies reached to 81.3%. For the practical anaerobic sludge digester liquor treatment, the average TN removal efficiency of 72% was obtained. A protein substance was shown to be the most abundant extracellular polymeric substances (EPS) in the granular sludge with almost two times more in the attached sludge of the Biofix reactor. Considering the EPS levels and observation by scanning electron microscopy, the anammox granules in the Biofix reactor were showing dense state. Results of DNA analyses indicated that the KSU-1 strain might prefer relatively low nutrient levels, while the KU2 strain might be better suited for the high media concentration. Other kinds of bacteria were also identified with the potentials for consuming the dissolved oxygen in the influent and facilitating anammox bacteria surviving under aerobic conditions. [ABSTRACT FROM AUTHOR]

This paper reports on the measurement of the thermal conductivity of an individual silicon carbide (SiC) nanowire of 140 nm diameter. T-type nanosensor and high-resolution transmission electron microscopy (HRTEM) are applied to obtain reliable property data. HRTEM images show that this nanowire has highly-crystalline SiC core of 126 nm diameter and surrounding amorphous silicon-dioxide layer of 7 nm thickness. Thermal contact resistance is estimated by using a simple analysis of the amorphous-carbon nanostructure between nanowire and nanosensor. Obtained apparent thermal conductivity of this nanowire suggests the thermal conductivity of SiC core is over 100W⋅m-1⋅K-1 at room temperature, which is much greater than the past-reported data of thin film but less than the pure bulk data. Compared with bulk samples, phonon scattering mechanism is also discussed. [ABSTRACT FROM AUTHOR]

Abstract: Paper sludge ash was partially converted into zeolites by reaction with 3M NaOH solution at 90°C for 24h. The paper sludge ash had a low abundance of Si and significant Ca content, due to the presence of calcite that was used as a paper filler. Diatomite was added to the NaOH solution to increase its Si content in order to synthesize zeolites with high cation exchange capacity. Diatomite residue was filtered from solution before addition of ash. The original ash without addition of diatomite yielded hydroxysodalite with a cation exchange capacity ca. 50cmol/kg. Addition of Si to the solution yielded Na-P1 (zeolite-P) with a higher cation exchange capacity (ca. 130cmol/kg). The observed concentrations of Si and Al in the solution during the reaction explain the crystallization of these two phases. The reaction products were tested for their capacity for PO4 3− removal from solution as a function of Ca2+ content, suggesting the formation of an insoluble Ca-phosphate salt. The product with Na-P1 exhibits the ability to remove NH4 + as well as PO4 3− from solution in concentrations sufficient for application in water purification. Both NH4 + and PO4 3− removal showed little variation with pH between 5 and 9. Alternative processing methods of zeolite synthesis, including the addition of ash to an unfiltered Si–NaOH solution and addition of a dry ash/diatomite mixture to NaOH solution, were tested. The third process yielded materials with lower cation exchange capacity due to formation of hydroxysodalite. The second process results in a product with relatively high cation exchange capacity, and reduces the number of processing steps necessary for zeolite synthesis. [Copyright &y& Elsevier]

Gordonia sp. strain P8219, a strain able to decompose di-2-ethylhexyl phthalate, was isolated from machine oil-contaminated soil. Mono-2-ethylhexyl phthalate hydrolase was purified from cell extracts of this strain. This enzyme was a 32,164-Da homodimeric protein, and it effectively hydrolyzed monophthalate esters, such as monoethyl, monobutyl, monohexyl, and mono-2-ethylhexyi phthalate. The Km and Vmax values for mono-2-ethylhexyl phthalate were 26.9 ± 4.3 μM and 18.1 ± 0.9 μmol/min · mg protein, respectively. The deduced amino acid sequence of the enzyme exhibited less than 30% homology with those of meta-cleavage hydrolases which are serine hydrolases but exhibited no significant homology with the sequences of serine esterases. The pentapeptide motif GXSXG, which is conserved in serine hydrolases, was present in the sequence. The enzymatic properties and features of the primary structure suggested that this enzyme is a novel enzyme belonging to an independent group of serine hydrolases. [ABSTRACT FROM AUTHOR]

Typically, the ash from incineration of paper sludge contains a high percentage of Ca in the form of anorthite (CaAl2Si2O8) and gehlenite (Ca2Al2SiO7). The Ca in the sludge originates from calcite that is included in paper as fillers. We applied acid leaching with HCl on the ash to reduce its Ca content. Zeolite was then synthesized from the leached ash through reaction with 2.5 M NaOH solution at 80 °C for 24 hours. The fraction of Ca and Al extracted from the ash correlates with the pH of the leachant. We determined the leachant pH (after 24 hours of leaching) associated with the Ca: Al:Si ratio in the leached ash that provided optimal production of zeolites with high cation-exchange capacity. During acid leaching, gehlenite dissolved out at higher pH than anorthite. In the case of pH > 5, both gehlenite and anorthite remained in the ash, and hydroxysodalite and LTA (Linde Type A) were synthesized in the product. In the case of pH = 1-5 in the leachant, gehlenite dissolved out but anorthite remained in the ash, and LTA and Na-P1 (zeolite-P) were produced. In the case of pH < 1, both gehlenite and anorthite dissolved out, and only Na-P1 was produced. The cation-exchange capacities of the products that contained hydroxysodalite, LTA, and Na-P1 were approximately 130, 200, and 120 cmol/kg, respectively. We conclude that acid leaching of paper sludge ash controls which of the three zeolite phases form, and that LTA and Na-Pl exhibit a higher cation-exchange capacity than hydroxysodalite. The most efficient production of zeolites with high cation-exchange capacity (about 220 cmol/kg) is obtained after leaching the sludge ash in solutions of around pH = 3. At this low pH, gehlenite has already dissolved out of the sludge ash, half the Ca content of the ash has been leached out, Si has not yet been leached, but Al has begun to be leached. After ash zeolitization, LTA coexists with Na-Pl in t... [ABSTRACT FROM AUTHOR]

In the fiscal year of 1998, The Ministry of Economy Trade and Industry (METI) of Japan launched a national 5-year project called the Humanoid Robotics Project. As a part of this project, we are developing a novel humanoid robot telexistence (tel-existence) system to assist and cooperate with people. This paper describes a newly developed telexistence cockpit for humanoid robot control, and shows a technical demonstration to evaluate the developed cockpit and the robot. A human operator controls the robot within the remote cockpit as if he or she were inside the robot itself. The telexistence cockpit consists of three subsystems: a three-dimensional (3D) audio/visual display subsystem, a telexistence master subsystem, and a communication subsystem between the cockpit and the robot. A series of real images are captured by cameras mounted on the robot and presented on the visual display, and the human operator in the cockpit observes them with a sensation of real-time presence. He or she can intuitively control the arms and hands of the slave robot through the telexistence master subsystem with force feedback. [ABSTRACT FROM AUTHOR]

A sensitive and simple high-performance liquid chromatography assay for benzalkonium chloride (BZK) in biological samples was developed. The biological samples, spiked with domiphen used as an internal standard, were purified by solid-phase extraction. The major homologues of BZK in pharmaceutical products (C12 and C14) were eluted at 24 and 36 min using a YMC-Pack CN column (4.6×250 mm, 5 μm) with a mobile phase, mixture of acetonitrile and sodium acetate buffer (48:52), and monitored at 254 nm. The most dominant component C12 was selected as an indicator to quantify BZK, and adjustment was then done based on the proportion of C12 in the BZK product. Good linearity was obtained in the range of 0.1–3 μg, and the limit of detection was 20 ng as a loaded amount on column. The recoveries of BZK in serum and tissues ranged from 54 to 90%. In a practical case, 0.16 μg/ml of BZK was quantified in serum collected several hours after accidental ingestion. The method is simple, sensitive and reliable for determining BZK levels in practical biological samples. [ABSTRACT FROM AUTHOR]

Objectives: The influence on enamel fluoride uptake of reducing the NaF concentration of a mouthrinse solution by half, from 0.05-0.025%, was examined in elementary school children. Methods: The participatory elementary school had employed school-based fluoride mouthrinse with 0.05% NaF solution since 1983, and in 1993, the NaF content was lowered to 0.025%. Dental examination and enamel biopsy of maxillary central incisors were performed just before the NaF content reduction for children who had received the daily 0.05% NaF mouthrinse for 2-5 years since 6 years of age. During 1993-95, dental examination and enamel biopsy were undertaken for newly enrolled 6-year-old children, and the same subjects were re-examined after 2 years. The same series of examinations were conducted for the control group, which consisted of children belonging to another elementary school without a fluoride program. Results: The enamel fluoride concentration of 8-year-old children who used the 0.025% NaF mouthrinse for 2 years was 1810±535 ppm, reaching the same order as that in chilclren who continued using the 0.05% NaF mouthrinse (1863±1358 ppm), and was significantly higher than that of children without fluoride programs (1113 ± 303 ppm). There was no considerable increase of mean DMFT values due to the NaF content reduction. ConcIusions: It was concluded that a daily 0.025% NaF mouthrinse provides the similar fluoride content for enamel as 0.05% NaF mouthrinse as long as the rinsing frequency is maintained. [ABSTRACT FROM AUTHOR]

• Rat HEV ORF3 has the PxYPMP motif in place of the PSAP motif for human HEV ORF3. • Rat HEV utilizes multivesicular body sorting and exosomal pathway for its release. • Association of host factors Tsg101, Alix and Vps4A/B with rat HEV egress was noted. • Unlike human HEV ORF3, rat HEV ORF3 did not bind to Tsg101. • A 90-kDa host protein that could be a candidate alternative to Tsg101 was found. Recent reports have shown that rat hepatitis E virus (HEV) is capable of infecting humans. We also successfully propagated rat HEV into human PLC/PRF/5 cells, raising the possibility of a similar mechanism shared by human HEV and rat HEV. Rat HEV has the proline-rich sequence, PxYPMP, in the open reading frame 3 (ORF3) protein that is indispensable for its release. However, the release mechanism remains unclear. The overexpression of dominant-negative (DN) mutant of vacuolar protein sorting (Vps)4A or Vps4B decreased rat HEV release to 23.9 % and 18.0 %, respectively. The release of rat HEV was decreased to 8.3 % in tumor susceptibility gene 101 (Tsg101)-depleted cells and to 31.5 % in apoptosis-linked gene 2-interacting protein X (Alix)-depleted cells. Although rat HEV ORF3 protein did not bind to Tsg101, we found a 90-kDa protein capable of binding to wild-type rat HEV ORF3 protein but not to ORF3 mutant with proline to leucine mutations in the PxYPMP motif. Rat HEV release was also decreased in Ras-associated binding 27A (Rab27A)- or hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs)-depleted cells (to 20.1 % and 18.5 %, respectively). In addition, the extracellular rat HEV levels in the infected PLC/PRF/5 cells were increased after treatment with Bafilomycin A1 and decreased after treatment with GW4869. These results indicate that rat HEV utilizes multivesicular body (MVB) sorting for its release and that the exosomal pathway is required for rat HEV egress. A host protein alternative to Tsg101 that can bind to rat HEV ORF3 should be explored in further study. [ABSTRACT FROM AUTHOR]

Cu-catalyzed 6π-electrocyclization of triene-substrates of type (I) provides a facile access to pyrido[4,3-b]carbazoles, which represent advanced intermediates for the construction of various ellipticine and olivacine alkaloids. [ABSTRACT FROM AUTHOR]

Graphene oxide (GO) is hydrophilic and swells significantly when in contact with water. Here, we investigate the change in thickness of multilayer graphene oxide membranes due to intercalation of water, via humidity-controlled observation in an environmental scanning electron microscope (ESEM). The thickness increases reproducibly with increasing relative humidity. Electron energy loss spectroscopy (EELS) reveals the existence of water ice under cryogenic conditions, even in high vacuum environment. Additionally, we demonstrate that freezing then thawing water trapped in the multilayer graphene oxide membrane leads to the opening up of micron-scale inter-lamellar voids due to the expansion of ice crystals. [ABSTRACT FROM AUTHOR]

The synthesis of alkaloid (I) is achieved in six steps in 60% overall yield from dihydroxymethylchromanone. [ABSTRACT FROM AUTHOR]