Physicochemical Properties, Cytocompatibility, and Biocompatibility of a Bioactive Glass Based Retrograde Filling Material.

The ideal retrograde filling material that is easy to handle, has good physicochemical properties, and is biocompatible has not yet been developed. The current study reports the development of a novel bioactive glass based powder for use as a retrograde filling material that is capable of altering the consistency and hardening rate of mixtures when mixed with existing bioactive glass based cement. Furthermore, its physicochemical properties, in vitro effects on human cementoblast-like cells, and in vivo effects on inflammatory responses were evaluated. The surface of the hardened cement showed the formation of hydroxyapatite-like precipitates and calcium and silicate ions were eluted from the cement when the pH level was stabilized at 10.5. Additionally, the cement was found to be insoluble and exhibited favorable handling properties. No adverse effects on viability, proliferation, and expression of differentiated markers were observed in the in vitro experiment, and the cement was capable of inducing calcium deposition in the cells. Moreover, the cement demonstrated a lower number of infiltrated inflammatory cells compared to the other materials used in the in vivo mouse subcutaneous implantation experiment. These findings suggest that the retrograde filling material composed of bioactive glass and the novel bioactive glass based powder exhibits favorable physicochemical properties, cytocompatibility, and biocompatibility.

Correlation between seed longevity and RNA integrity in the embryos of rice seeds.

The mature embryos of rice seeds contain translatable mRNAs required for the initial phase of germination. To clarify the relationship between seed longevity and RNA integrity in embryos, germinability and stability of embryonic RNAs were analyzed using the seeds of japonica rice cultivars subjected to controlled deterioration treatment (CDT) or long periods of storage. Degradation of RNA from embryos of a japonica rice cultivar "Nipponbare" was induced by CDT before the decline of the germination rate and we observed a positive relationship between seed germinability and integrity of embryonic RNAs. Moreover, this relationship was confirmed in the experiments using aged seeds from the "Nipponbare", "Sasanishiki" and "Koshihikari" rice cultivars. In addition, the RNA integrity number (RIN) values, calculated using electrophoresis data and Agilent Bioanalyzer software, had a positive correlation with germinability (R2=0.75). Therefore, the stability of embryonic RNAs required for germination is involved in maintaining seed longevity over time and RIN values can serve as a quantitative indicator to evaluate germinability in rice.

Effect of Bioactive Glass-Based Root Canal Sealer on the Incidence of Postoperative Pain after Root Canal Obturation.

The purpose of this study is to evaluate the effect of a bioactive glass-based root canal sealer, Nishika Canal Sealer BG (CS-BG), on the incidence of postoperative pain (PP) after root canal obturation (RCO). Eleven dentists performed pulpectomy or infected root canal treatments for 555 teeth. During RCO, CS-BG was used. After RCO, the rate of PP and the factors affecting PP (pain during RCO and pain immediately after RCO) were analyzed. PP was observed in eight teeth (1.5%), and within 7 days after RCO, there were no teeth with pain. In these teeth with PP, there was a significant difference in the occurrence of pain during RCO, but not in the occurrence of pain immediately after RCO, when compared with pulpectomy and infected root canal treatment. These clinical results show that CS-BG has an excellent biocompatibility, and can suppress the distress of patients during RCO.

Selective Hydrogenation of Cinnamaldehyde over the Stepped and Plane Surface of Pd Nanoparticles with Controlled Morphologies by CO Chemisorption.

Carbon monoxide (CO) molecules are attracting attention as capping agents that control the structure of metal nanoparticles. In this study, we aimed to control the shape and surface structure of Pd particles by reducing the supported Pd precursor with CO. The reduction of Pd nanoparticles with CO promoted the exposure of step sites and generated spherical and concave-tetrahedral Pd particles on carbon and SiO2 supports. On the other hand, conventional H2-reduced Pd particles show a flattened shape. The preferential exposure of the step sites by the adsorbed CO molecules was supported by the density functional theory-calculated surface energy and the Wulff construction. Morphology- and surface-controlled Pd nanoparticles were used to study the surface structure and morphology effects of Pd nanoparticles on cinnamaldehyde (CAL) hydrogenation. With an increase in the fraction of step sites on Pd nanoparticles, the hydrogenation activity and selectivity of hydrocinnamaldehyde (HCAL) increased. On step sites, the adsorption of the C═C bond of CAL proceeded preferentially, and HCAL was efficiently and selectively generated.

Enhancement of Alkaline Hydrogen Oxidation Reaction of Ru-Ir Alloy Nanoparticles through Bifunctional Mechanism on Ru-Ir Pair Site.

Anion exchange membrane fuel cells (AEMFCs) are being developed for practical use. However, it is necessary to improve the hydrogen oxidation reaction (HOR) under alkaline conditions to enhance the performance of AEMFCs. In this study, carbon-supported Ru-Ir alloy nanoparticle catalysts (Ru-Ir/C) were developed because they offer higher HOR activity compared with the Pt-based catalysts. The mechanism of HOR enhancement on Ru-Ir/C was studied to reveal the effect of the surface composition of Ru/Ir on the HOR activity. The results showed that the HOR activity is related to the surface pair probability of Ru-Ir, Ru-Ru, and Ir-Ir but not to the hydrogen binding energy (HBE). In addition, the Ru-Ir pair site was found to be highly active, which can promote the HOR through a bifunctional mechanism that involves Ru-Ir pairs providing reactive OH- and H species, respectively.

Identification of active sites in CO oxidation over a Pd/Al2O3 catalyst.

The active sites of Pd/Al2O3 catalysts for CO oxidations were identified by investigating the dependence of CO oxidation activities on the surface structure and morphology of Pd nanoparticles. The maximum catalytic activity was obtained for Pd particles approximately 2 nm in particle size. We performed structural analyses on the Pd surface through infrared (IR) spectroscopy of the adsorbed CO molecules. A positive correlation was obtained between catalytic activity and the fraction of linear CO adsorbed on Pd corner sites and Pd(111) facets, indicating that these sites are highly active for CO oxidation. X-ray absorption fine structure (XAFS) and spherical aberration-corrected scanning transmission electron microscopy (Cs-STEM) measurements demonstrated that Pd nanoparticles less than 2 nm in particle size with amorphous-like structures and Pd particles with large, well-ordered structures favor the formation of a high fraction of corner sites and Pd(111) facets, respectively.

A rice gene that confers broad-spectrum resistance to ß-triketone herbicides.

The genetic variation of rice cultivars provides a resource for further varietal improvement through breeding. Some rice varieties are sensitive to benzobicyclon (BBC), a ß-triketone herbicide that inhibits 4-hydroxyphenylpyruvate dioxygenase (HPPD). Here we identify a rice gene, HIS1 (HPPD INHIBITOR SENSITIVE 1), that confers resistance to BBC and other ß-triketone herbicides. We show that HIS1 encodes an Fe(II)/2-oxoglutarate-dependent oxygenase that detoxifies ß-triketone herbicides by catalyzing their hydroxylation. Genealogy analysis revealed that BBC-sensitive rice variants inherited a dysfunctional his1 allele from an indica rice variety. Forced expression of HIS1 in Arabidopsis conferred resistance not only to BBC but also to four additional ß-triketone herbicides. HIS1 may prove useful for breeding herbicide-resistant crops.

Thermo-responsive allele of sucrose synthase 3 (Sus3) provides high-temperature tolerance during the ripening stage in rice (Oryza sativa L.).

High-temperature stress during the ripening stage leads to quality deterioration due to an increase in chalky grains in brown rice (Oryza sativa L.). In a previous study, we identified a QTL for Appearance quality of brown rice 1 (Apq1) using chromosome segment substitution lines of the indica cultivar 'Habataki' in the japonica cultivar 'Koshihikari' background and narrowed down the locus to a 48-kb region on chromosome 7. To verify the function and mechanisms of this QTL in grain appearance, in this study, we fine-mapped the gene and conducted high-temperature tolerance tests. As a result of the genetic mapping, we narrowed down the candidate region of Apq1 to a 19.4-kb region including three predicted genes. Among these, the temporal expression pattern of sucrose synthase 3 (Sus3) corresponded well with the high temperature-sensitive period during ripening, and expression of the 'Habataki' allele of Sus3 was increased under high-temperature condition. In addition, we transformed the 'Habataki' Sus3 gene into 'Nipponbare', and the transformants obtained high-temperature tolerance. Therefore, we conclude that the causal gene underlying the QTL Apq1 is the thermo-responsive Sus3 allele, and the increase in Sus3 expression under high-temperature condition during ripening leads to high-temperature tolerance in rice.

Identification of OsGGR2, a second geranylgeranyl reductase involved in α-tocopherol synthesis in rice.

Tocopherol (Toc) and tocotrienol (T3) are abundant in rice bran. Geranylgeranyl reductase (GGR) is an essential enzyme for Toc production that catalyzes the reduction of geranylgeranyl pyrophosphate and geranylgeranyl-chlorophyll. However, we found that a rice mutant line with inactivated Os02g0744900 (OsGGR1/LYL1/OsChl P) gene produces Toc, suggesting that rice plants may carry another enzyme with GGR activity. Using an RNA-mediated interference technique, we demonstrated that the Os01g0265000 ("OsGGR2") gene product has GGR activity. This result supports the existence of two GGR genes (OsGGR1 and OsGGR2) in rice, in contrast to Arabidopsis thaliana (thale cress) and cyanobacterium Synechocystis that each have only one GGR gene. We also produced rice callus with inactivated OsGGR1 and OsGGR2 that produced T3 but not Toc. Such rice callus could be used as a resource for production of pure T3 for nutraceutical applications.

The Metal-Support Interaction Concerning the Particle Size Effect of Pd/Al2 O3 on Methane Combustion.

The particle size effect of Pd nanoparticles supported on alumina with various crystalline phases on methane combustion was investigated. Pd/θ, α-Al2 O3 with weak metal-support interaction showed a volcano-shaped dependence of the catalytic activity on the size of Pd particles, and the catalytic activity of the strongly interacted Pd/γ-Al2 O3 increased with the particle size. Based on a structural analysis of Pd nanoparticles using CO adsorption IR spectroscopy and spherical aberration-corrected scanning/transmission electron microscopy, the dependence of catalytic activity on Pd particle size and the alumina crystalline phase was due to the fraction of step sites on Pd particle surface. The difference in fraction of the step site is derived from the particle shape, which varies not only with Pd particle size but also with the strength of metal-support interaction. Therefore, this interaction perturbs the particle size effect of Pd/Al2 O3 for methane combustion.

Identification of a Genetic Factor Required for High γ-Isoform Concentration in Rice Vitamin E.

The γ-isoforms of tocopherols (Tc) and tocotrienols (T3) possess high biological activities in comparison to the α-isoforms. The concentrations of Tc and T3 isoforms in rice (Oriza sativa) was cultivar-dependent. Using chromosome segment substitution lines (CSSLs) and near isogenic lines (NILs) of indica cultivar "Kasalath" in a japonica cultivar "Koshihikari" genetic background, the Kasalath genomic segment on chromosome 2 was determined to be responsible for the high γ-isoform concentration: γ-tocopherol methyltransferase (γ-TMT) was identified as a candidate gene. An amino acid substitution in the coding region and several nucleotide polymorphisms, including an insertion of 10 base pairs in the promoter region, were identified. Gene expression analysis revealed that low expression levels of the γ-TMT gene in Kasalath were not associated with the γ-isoform concentration. Genetic variations in the coding region of the γ-TMT gene may play a major role in determining the γ-isoform concentration. This information could be used to breed rice with a high γ-isoform content.

Kinetic study of the quenching reaction of singlet oxygen by seven rice bran extracts in ethanol solution. Development of a singlet oxygen absorption capacity (SOAC) assay method.

Measurements of singlet oxygen ((1)O2) quenching rates (kQ (S)) and the relative singlet oxygen absorption capacity (SOAC) values were performed for seven rice bran extracts 1-7, which contained different concentrations of antioxidants (AOs) (such as α-, ß-, γ-, and δ-tocopherols and -tocotrienols, three carotenoids (lutein, ß-carotene, and zeaxanthin), and γ-oryzanol), in ethanol at 35 °C using UV-vis spectrophotometry. The concentrations of four tocopherols and four tocotrienols, three carotenoids, and γ-oryzanol contained in the extracts were determined using HPLC-MS/MS, UV-HPLC, and UV-vis absorption spectroscopy, respectively. Furthermore, comparisons of kQ (S) (Obsd.) values observed for the above extracts 1-7 with the sum of the product {[Formula: see text] [AO-i]} of the [Formula: see text] values obtained for each AO-i and the concentration ([AO-i]) of AO-i contained in extracts 1-7 were performed. From the results, it has been ascertained that the SOAC method is applicable to general food extracts to evaluate their (1)O2-quenching activity.

Accumulation of long-lived mRNAs associated with germination in embryos during seed development of rice.

Mature dry seeds contain translatable mRNAs called long-lived mRNAs. Early studies have shown that protein synthesis during the initial phase of seed germination occurs from long-lived mRNAs, without de novo transcription. However, the gene expression systems that generate long-lived mRNAs in seeds are not well understood. To examine the accumulation of long-lived mRNAs in developing rice embryos, germination tests using the transcriptional inhibitor actinomycin D (Act D) were performed with the Japonica rice cultivar Nipponbare. Although over 70% of embryos at 10 days after flowering (DAF) germinated in the absence of the inhibitor, germination was remarkably impaired in embryos treated with Act D. In contrast, more than 70% of embryos at 20, 25, 30 and 40 DAF germinated in the presence of Act D. The same results were obtained when another cultivar, Koshihikari, was used, indicating that the long-lived mRNAs required for germination predominantly accumulate in embryos between 10 and 20 DAF during seed development. RNA-Seq identified 529 long-lived mRNA candidates, encoding proteins such as ABA, calcium ion and phospholipid signalling-related proteins, and HSP DNA J, increased from 10 to 20 DAF and were highly abundant in 40 DAF embryos of Nipponbare and Koshihikari. We also revealed that these long-lived mRNA candidates are clearly up-regulated in 10 DAF germinating embryos after imbibition, suggesting that the accumulation of these mRNAs in embryos is indispensable for the induction of germination. The findings presented here may facilitate in overcoming irregular seed germination or producing more vigorous seedlings.

Kinetic study of the scavenging reaction of the aroxyl radical by seven kinds of rice bran extracts in ethanol solution. Development of an aroxyl radical absorption capacity (ARAC) assay method.

Recently, a new assay method that can quantify the aroxyl radical (ArO•) absorption capacity (ARAC) of antioxidants (AOHs) was proposed. In the present work, the second-order rate constants (ks(Extract)) and ARAC values for the reaction of ArO• with seven kinds of rice bran extracts 1-7, which contain different concentrations of α-, ß-, γ-, and δ-tocopherols and -tocotrienols (α-, ß-, γ-, and δ-Tocs and -Toc-3s) and γ-oryzanol, were measured in ethanol at 25 °C using stopped-flow spectrophotometry. The ks(Extract) value (1.26 × 10(-2) M(-1) s(-1)) of Nipponbare (extract 1) with the highest activity was 1.5 times larger than that (8.29 × 10(-3)) of Milyang-23 (extract 7) with the lowest activity. The concentrations (in mg/100 g) of α-, ß-, γ-, and δ-Tocs and -Toc-3s and γ-oryzanol found in the seven extracts 1-7 were determined using HPLC-MS/MS and UV-vis absorption spectroscopy, respectively. From the results, it has been clarified that the ArO•-scavenging rates (ks(Extract)) (that is, the relative ARAC value) obtained for the seven extracts 1-7 may be approximately explained as the sum of the product {Σ ks(AOH-i) [AOH-i]/10(5)} of the rate constant (ks(AOH-i)) and the concentration ([AOH-i]/10(5)) of AOH-i (Tocs, Toc-3s, and γ-oryzanol) included in rice bran extracts. The contribution of γ-oryzanol to the ks(Extract) value was estimated to be between 3.0-4.7% for each extract. Taken together, these results suggest that the ARAC assay method is applicable to general food extracts.

Genetic dissection of black grain rice by the development of a near isogenic line.

Rice (Oryza sativa L.) can produce black grains as well as white. In black rice, the pericarp of the grain accumulates anthocyanin, which has antioxidant activity and is beneficial to human health. We developed a black rice introgression line in the genetic background of Oryza sativa L. 'Koshihikari', which is a leading variety in Japan. We used Oryza sativa L. 'Hong Xie Nuo' as the donor parent and backcrossed with 'Koshihikari' four times, resulting in a near isogenic line (NIL) for black grains. A whole genome survey of the introgression line using DNA markers suggested that three regions, on chromosomes 1, 3 and 4 are associated with black pigmentation. The locus on chromosome 3 has not been identified previously. A mapping analysis with 546 F2 plants derived from a cross between the black rice NIL and 'Koshihikari' was evaluated. The results indicated that all three loci are essential for black pigmentation. We named these loci Kala1, Kala3 and Kala4. The black rice NIL was evaluated for eating quality and general agronomic traits. The eating quality was greatly superior to that of 'Okunomurasaki', an existing black rice variety. The isogenicity of the black rice NIL to 'Koshihikari' was very high.

Introgression of two chromosome regions for leaf photosynthesis from an indica rice into the genetic background of a japonica rice.

Increases in rates of individual leaf photosynthesis (P n) are critical for future increases of rice yields. A previous study, using introgression lines derived from a cross between indica cultivar Habataki, with one of the highest recorded values of P n, and the Japanese elite cultivar Koshihikari, identified four QTLs (qCAR4, qCAR5, qCAR8, and qCAR11) that affect P n. The present study examined the combined effect of qCAR4 and qCAR8 on P n in the genetic background of Koshihikari. The pyramided near-isogenic line NIL(qCAR4+qCAR8) showed higher P n than both NIL(qCAR4) and NIL(qCAR8), equivalent to that of Habataki despite being due to only two out of the four QTLs. The high P n of NIL(qCAR4+qCAR8) may be attributable to the high leaf nitrogen content, which may have been inherited from NIL(qCAR4), to the large hydraulic conductance due to the large root surface area from NIL(qCAR4), and to the high hydraulic conductivity from NIL(qCAR8). It might be also attributable to high mesophyll conductance, which may have been inherited from NIL(qCAR4). The induction of mesophyll conductance and the high leaf nitrogen content and high hydraulic conductivity could not be explained in isolation from the Koshihikari background. These results suggest that QTL pyramiding is a useful approach in rice breeding aimed at increasing P n.

Identification of a novel gene (Apq1) from the indica rice cultivar 'Habataki' that improves the quality of grains produced under high temperature stress.

The appearance of brown rice grown under high temperature conditions is an important characteristic for improvement in Japanese rice breeding programs. We performed a QTL analysis of the appearance quality of brown rice using chromosome segment substitution lines of the indica cultivar 'Habataki' in the 'Koshihikari' genetic background. A line carrying a 'Habataki' segment on chromosome 7 showed a high percentage of perfect grains produced under high temperature conditions during the ripening period. To verify the role of this segment, and to narrow down the region containing the useful allele, substitution mapping was performed using multiple paired lines. As a result, the chromosomal location of a gene that we named Appearance quality of brown rice 1 (Apq1) was delimited to a 48-kb region. In addition, we developed an Apq1-near isogenic line (NIL) to evaluate the effect of Apq1 on various agronomic traits. Under high temperature conditions during the ripening period, the Apq1-NIL produced significantly higher percentages of perfect grains than 'Koshihikari'. Other agronomic traits, including yield and palatability, were similar between the Apq1-NIL and 'Koshihikari'. Therefore, the 'Habataki' allele of Apq1 will be useful in breeding programs aimed at improving the quality of grains ripened under high temperature conditions.

Investigation of tocotrienol biosynthesis in rice (Oryza sativa L.).

Rice tocotrienol (T3) has gained attention due to its physiological activities (e.g., antiangiogenesis). However, the biosynthetic pathway for T3 production in rice grain has not been well studied. We hypothesized that T3 biosynthesis enzymes and/or precursors play an important role in T3 production in whole grain. This proposal was evaluated in rice (Oryza sativa L.) by PCR and HPLC techniques. Grain tocopherol as well as flag leaf vitamin E levels were also investigated for comparison. For rice samples 14 days after flowering, grain was abundant in T3, but not in flag leaf. Expression of a gene encoding homogentisate geranylgeranyltransferase (HGGT, which has long been believed to be important for T3 production) differed significantly between grain and flag leaf. We then investigated rice samples during the grain maturation period, and found that grain T3 and HGGT levels increased in the early stage and then reached a plateau. T3 precursors such as homogentisate and geranylgeranyl pyrophosphate decreased during maturation. No increase in grain T3 from the middle to late stages of maturation and a decrease in T3 precursors during maturation suggest that HGGT would be an essential, but not limiting factor for T3 biosynthesis, and T3 precursors could regulate the T3 level in grain. The results of this study would be useful for nutraceutical purposes (e.g., development of T3-overproducing rice for the prevention of angiogenic disorders).

Identification and characterization of genomic regions on chromosomes 4 and 8 that control the rate of photosynthesis in rice leaves.

DNA marker-assisted selection appears to be a promising strategy for improving rates of leaf photosynthesis in rice. The rate of leaf photosynthesis was significantly higher in a high-yielding indica variety, Habataki, than in the most popular Japanese variety, Koshihikari, at the full heading stage as a result of the higher level of leaf nitrogen at the same rate of application of nitrogen and the higher stomatal conductance even when the respective levels of leaf nitrogen were the same. The higher leaf nitrogen content of Habataki was caused by the greater accumulation of nitrogen by plants. The higher stomatal conductance of Habataki was caused by the higher hydraulic conductance. Using progeny populations and selected lines derived from a cross between Koshihikari and Habataki, it was possible to identify the genomic regions responsible for the rate of photosynthesis within a 2.1 Mb region between RM17459 and RM17552 and within a 1.2 Mb region between RM6999 and RM22529 on the long arm of chromosome 4 and on the short arm of chromosome 8, respectively. The designated region on chromosome 4 of Habataki was responsible for both the increase in the nitrogen content of leaves and hydraulic conductance in the plant by increasing the root surface area. The designated region on chromosome 8 of Habataki was responsible for the increase in hydraulic conductance by increasing the root hydraulic conductivity. The results suggest that it may be possible to improve photosynthesis in rice leaves by marker-assisted selection that focuses on these regions of chromosomes 4 and 8.

New approach for rice improvement using a pleiotropic QTL gene for lodging resistance and yield.

The use of fertilizer results in tall rice plants that are susceptible to lodging and results in reduced plant yields. In this study, using chromosome segment substitution lines, we identified an effective quantitative trait loci (QTL) for culm strength, which was designated STRONG CULM2 (SCM2). Positional cloning of the gene revealed that SCM2 was identical to ABERRANT PANICLE ORGANIZATION1 (APO1), a gene previously reported to control panicle structure. A near-isogenic line carrying SCM2 showed enhanced culm strength and increased spikelet number because of the pleiotropic effects of the gene. Although SCM2 is a gain-of-function mutant of APO1, it does not have the negative effects reported for APO1 overexpression mutants, such as decreased panicle number and abnormal spikelet morphology. The identification of lodging-resistance genes by QTL analysis combined with positional cloning is a useful approach for improving lodging resistance and overall productivity in rice.