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Silicon (Si) is important for stable and high yields in rice (Oryza sativa), a typical Si hyperaccumulator. The high Si accumulation is achieved by the cooperation of 2 Si transporters, LOW SILICON 1 (OsLsi1) and OsLsi2, which are polarly localized in cells of the root exodermis and endodermis. However, the mechanism underlying their polar localization is unknown. Here, we identified amino acid residues critical for the polar localization of OsLsi1. Deletion of both N- and C-terminal regions resulted in the loss of its polar localization. Furthermore, the deletion of the C-terminus inhibited its trafficking from the endoplasmic reticulum to the plasma membrane. Detailed site-directed mutagenesis analysis showed that Ile18 at the N-terminal region and Ile285 at the C-terminal region were essential for the polar localization of OsLsi1. Moreover, a cluster of positively charged residues at the C-terminal region is also required for polar localization. Phosphorylation and Lys modifications of OsLsi1 are unlikely to be involved in its polar localization. Finally, we showed that the polar localization of OsLsi1 is required for the efficient uptake of Si. Our study not only identified critical residues required for the polar localization of OsLsi1, but also provided experimental evidence for the importance of transporter polarity for efficient nutrient uptake.

Rice is able to accumulate high concentrations of silicon (Si) in the shoots, and this ability is required for the mitigation of abiotic and biotic stresses. Although transporters for Si uptake have been identified, a transporter for the xylem loading of Si has not been found. We functionally characterized a Si transporter, OsLsi3, in terms of tissue-specific localization, knockout line phenotype and mathematic simulation. OsLsi3 was shown to be an efflux Si transporter. OsLsi3 was mainly expressed in the mature root region, and its expression was downregulated by Si. Immunostaining with a specific antibody showed that OsLsi3 was localized to the pericycle in the roots, without polarity. However, when it was expressed under the control of the OsLsi2 promoter, OsLsi3 became polarly localized to the proximal side of both the exodermis and endodermis. Knockout of this gene resulted in decreased Si uptake and concentration in the xylem sap under low Si supply, but not under high Si supply. Mathematical modeling showed that localization of OsLsi3 to the pericycle accounts for c. 30% of the total Si loading to the xylem under low Si concentrations. In summary, OsLsi3 was involved in the xylem loading of Si in rice roots, which is required for the efficient root-to-shoot translocation of Si.

Ammonium is a preferential nitrogen form for rice (Oryza sativa) grown in paddy field, but the molecular mechanisms for ammonium uptake have not been well understood. We functionally characterized three members belonging to ammonium transporter 1 (AMT1) and investigated their contributions to ammonium uptake. Spatial expression analysis showed that the upregulated expression of OsAMT1;1 and OsAMT1;2 and downregulated expression of OsAMT1;3 by ammonium were higher in the root mature region than in the root tips. All OsAMT1 members were polarly localized at the distal side of exodermis in the mature region of crown roots and lateral roots. Upon exposure to ammonium, localization of OsAMT1;1 and OsAMT1;2 was also observed in the endoplasmic reticulum, but their abundance in the plasma membrane was not changed. Single knockout of either gene did not affect ammonium uptake, but knockout of all three genes resulted in 95% reduction of ammonium uptake. However, the nitrogen uptake did not differ between the wild-type rice and triple mutants at high ammonium and nitrate supply. Our results indicate that three OsAMT1 members are cooperatively required for uptake of low ammonium in rice roots and that they undergo a distinct regulatory mechanism in response to ammonium.

Purpose: Continuous positive airway pressure (CPAP) therapy is a well-established treatment for obstructive sleep apnea (OSA). In this study, we aimed to assess the effects of CPAP therapy on nocturnal blood pressure (NBP) fluctuations (NBPFs) during rapid eye movement (REM) and non-REM sleep, and the NBPF patterns in patients with OSA. Methods: This retrospective study included 34 patients with moderate-to-severe OSA who underwent polysomnography using pulse transit time pre-CPAP therapy and 3–6 months post-CPAP therapy. We investigated NBP and the frequency of NBPFs in REM and non-REM sleep, as well as changes in NBPFpatterns post-CPAP therapy. Results: The apnea-hypopnea index (AHI), arousal index, systolic NBP, diastolic NBP, diurnal systolic BP (SBP), diastolic BP (DBP), and frequency of NBPFs in REM and non-REM sleep were significantly reduced post-CPAP therapy (all p < 0.01). A higher pre-CPAP AHI was associated with a reduction in diurnal SBP, systolic NBP, and NBPFs post-CPAP (r = 0.32, p = 0.028; r = 0.40, p = 0.019; and r = 0.51, p = 0.002; respectively). However, 58.8% of the patients showed no NBPF pattern change with CPAP therapy. Conclusion: In REM and non-REM sleep, almost all sleep-related parameters, NBP, and frequency of NBPFs significantly improved post-CPAP therapy, whereas NBPF patterns showed varying changes 3–6 months post-CPAP therapy. These results suggest that factors other than OSA influence NBPF pattern changes.

Rice is a major dietary source of the toxic metal cadmium (Cd), and reducing its accumulation in the grain is therefore important for food safety. We selected two cultivars with contrasting Cd accumulation and generated transgenic lines overexpressing OsNRAMP5, which encodes a major influx transporter for manganese (Mn) and Cd. We used two different promoters to control the expression, namely OsActin1 and maize Ubiquitin. Overexpression of OsNRAMP5 increased Cd and Mn uptake into the roots, but markedly decreased Cd accumulation in the shoots, whilst having a relatively small effect on Mn accumulation in the shoots. The overexpressed OsNRAMP5 protein was localized to the plasma membrane of all cell types in the root tips and lateral root primordia without polarity. Synchrotron X-ray fluorescence mapping showed that the overexpression lines accumulated more Cd in the root tips and lateral root primordia compared with the wild-type. When grown in three Cd-contaminated paddy soils, overexpression of OsNRAMP5 decreased concentration of Cd in the grain by 49–94% compared with the wild type. OsNRAMP5-overexpression plants had decreased Cd translocation from roots to shoots as a result of disruption of its radial transport into the stele for xylem loading, demonstrating the effect of transporter localization and polarity on ion homeostasis.

Silicon (Si) is an important nutrient required for sustainable and high production of rice and its uptake is mediated by a pair of influx (OsLsi1)–efflux (OsLsi2) transporters showing polar localization. However, the mechanisms underlying their polarity are unknown. Here, we revealed that the polarity of the Si transporters depends on cell types. The polar localization of both OsLsi1 and OsLsi2 was not altered by Si supply, but their protein abundance was reduced. Double immunostaining showed that localization of OsLsi1 and OsLsi2 was separated at the edge of the lateral polar domain by Casparian strips in the endodermis, whereas they were slightly overlapped at the transversal side of the exodermis. When OsLsi1 was ectopically expressed in the shoots, it showed polar localization at the xylem parenchyma cells of the basal node and leaf sheath, but not at the phloem companion cells. Ectopic expression of non-polar Si transporters, barley HvLsi2 and maize ZmLsi2 in rice, resulted in their polar localization at the proximal side. The polar localization of OsLsi1 and OsLsi2 was not altered by inhibition of clathrin-mediated endocytosis (CME) by dominant-negative induction of dynamin-related protein1A and knockout of mu subunit of adaptor protein 2 complex, although the knockout mutants of OsAP2M gene showed dwarf phenotype. These results indicate that CME is not required for the polar localization of Si transporters. Taken together, our results indicate that CME-independent machinery controls the polar localization of Si transporters in exodermis, endodermis of root cells and xylem parenchyma cells.

Uptake of boron (B) in rice (Oryza sativa) is mediated by the Low silicon rice 1 (OsLsi1) channel, belonging to the NOD26-like intrinsic protein III subgroup, and the efflux transporter B transporter 1 (OsBOR1). However, it is unknown how these transporters cooperate for B uptake and how they are regulated in response to B fluctuations. Here, we examined the response of these two transporters to environmental B changes at the transcriptional and posttranslational level. OsBOR1 showed polar localization at the proximal side of both the exodermis and endodermis of mature root region, forming an efficient uptake system with OsLsi1 polarly localized at the distal side of the same cell layers. Expression of OsBOR1 and OsLsi1 was unaffected by B deficiency and excess. However, although OsLsi1 protein did not respond to high B at the protein level, OsBOR1 was degraded in response to high B within hours, which was accompanied with a significant decrease of total B uptake. The high B-induced degradation of OsBOR1 was inhibited in the presence of MG-132, a proteasome inhibitor, without disturbance of the polar localization. In contrast, neither the high B-induced degradation of OsBOR1 nor its polarity was affected by induced expression of dominant-negative mutated dynamin-related protein 1A (OsDRP1AK47A) or knockout of the mu subunit (AP2M) of adaptor protein-2 complex, suggesting that clathrin-mediated endocytosis is not involved in OsBOR1 degradation and polar localization. These results indicate that, in contrast to Arabidopsis thaliana, rice has a distinct regulatory mechanism for B uptake through clathrin-independent degradation of OsBOR1 in response to high B.

Rapid eye movement-related obstructive sleep apnea (REM-related OSA) is a polysomnographic phenotype. Nocturnal blood pressure (BP) fluctuations remain unclear in patients with REM-related OSA. We studied 27 patients with REM-related OSA, categorized as having REM-apnea-hypopnea index (REM-AHI) ≥ 5/h, REM-AHI/non-REM-AHI ≥ 2, and non-REM-AHI &lt, 15/h. Beat-to-beat systolic BP (SBP) variability and nocturnal SBP fluctuation patterns using pulse transit time (PTT) were investigated. The maximum increase and average nocturnal SBP were significantly higher in males than in females (p = 0.003 and p = 0.008, respectively). The rate of non-dipping patterns in nocturnal SBP fluctuations was 63% in all patients (males, 70%, females, 50%). Epworth Sleepiness Scale (ESS) and Self-rating Depression Scale (SDS) scores in females were higher than those in males (8.4 ± 6.1 vs. 13.4 ± 5.4 points, p = 0.04, 43.8 ± 7.9 vs. 52 ± 11.6 points, p = 0.04, respectively). A high proportion of patients with REM-related OSA had a non-dipping pattern. Using PPT, we observed that in patients with REM-related OSA, SBP variability was greater in males. Despite clinical symptoms being slightly more severe in females, nocturnal SBP fluctuations should be considered in male patients with REM-related OSA.

Nitrogen is one of the most important elements for plant growth, and urea is one of the most frequently used nitrogen fertilizers worldwide. Besides the exogenously-supplied urea to the soil, urea is endogenously synthesized during secondary nitrogen metabolism. Here, we investigated the contribution of a urea transporter, DUR3, to rice production using a reverse genetic approach combined with localization studies. Tos17 insertion lines for DUR3 showed a 50% yield reduction in hydroponic culture, and a 26.2% yield reduction in a paddy field, because of decreased grain filling. Because shoot biomass production and shoot total N was not reduced, insertion lines were disordered not only in nitrogen acquisition but also in nitrogen allocation. During seed development, DUR3 insertion lines accumulated nitrogen in leaves and could not sufficiently develop their panicles, although shoot and root dry weights were not significantly different from the wild-type. The urea concentration in old leaf harvested from DUR3 insertion lines was lower than that in wild-type. DUR3 promoter-dependent β-glucuronidase (GUS) activity was localized in vascular tissue and the midribs of old leaves. These results indicate that DUR3 contributes to nitrogen translocation and rice yield under nitrogen-deficient and field conditions.

Purpose Rapid eye movement (REM) obstructive sleep apnea (OSA) is a prevalent clinical phenotype. However, the literature focusing on the pathophysiology of REM OSA is limited. This study compared the proportion of individuals with a low respiratory arousal threshold between patients with REM and non-REM OSA. Methods REM OSA was defined as having an apnea–hypopnea index (AHI) ≥ 5 and AHI during REM (AHI-REM)/AHI during NREM (AHI-NREM) ≥ 2. REM OSA was sub-divided into REM-predominant OSA and REM-isolated OSA. REM-predominant OSA was defined as satisfying the definition of REM OSA and having an AHI-NREM ≥ 5. REM-isolated OSA was defined as satisfying the definition of REM OSA and having an AHI-NREM 58.3%, and nadir SpO2 > 82.5%. Results The proportions of individuals with low respiratory arousal thresholds among individuals with REM-predominant OSA and REM-isolated OSA were significantly higher (77.2% and 93.7%, respectively) than that of patients with non-REM OSA (48.6%). This was also true when the analysis was performed according to sex. Conclusion These results indicate that a low respiratory arousal threshold might be an important endotype that contributes to the pathogenesis of REM OSA, especially in REM-isolated OSA.

Rapid eye movement (REM)-related obstructive sleep apnea (OSA), a polysomnographic phenotype that affects 12&ndash, 36% of OSA patients, is defined by apnea and hypopnea events that predominantly or exclusively occur during REM sleep. Recent studies indicated that REM-related OSA was associated with the development of nocturnal non-dipping of systolic and diastolic blood pressure, metabolic syndrome, diabetes, and depressive symptoms. However, to date, the association between REM-related OSA and insomnia still remains unclear. We investigated whether there was a difference between REM- and non-REM-related OSA in terms of insomnia-related sleep disturbance as measured by the Pittsburgh Sleep Quality Index (PSQI) in 1736 patients with OSA. REM-related OSA showed a significant association with increased PSQI in all adjusted models. In the subgroup analysis, the coefficients of all models were higher in female than in male patients with REM-related OSA. Insomnia should be considered an important complaint in patients with REM-related OSA, and its indicators, such as the PSQI, should be included in routine diagnostic testing.

School attendance is essential for both academic and socialemotional development. However, increasing numbers of teenagers visit our sleep clinic because of school absenteeism caused by difficulty getting up or excessive morning sleepiness..

Rice is a major dietary source of the toxic metalloid arsenic. Reducing arsenic accumulation in rice grain is important for food safety. We generated transgenic rice overexpressing two aquaporin genes, OsNIP1;1 and OsNIP3;3, under the control of a maize ubiquitin promoter or the rice OsLsi1 promoter, and tested the effect on arsenite uptake and translocation. OsNIP1;1 and OsNIP3;3 were highly permeable to arsenite in Xenopus oocyte assays. Both transporters were localized at the plasma membrane. Knockout of either gene had little effect on arsenite uptake or translocation. Overexpression of OsNIP1;1 or OsNIP3;3 in rice did not affect arsenite uptake but decreased root-to-shoot translocation of arsenite and shoot arsenic concentration markedly. The overexpressed OsNIP1;1 and OsNIP3;3 proteins were localized in all root cells without polarity. Expression of OsNIP1;1 driven by the OsLsi1 promoter produced similar effects. When grown in two arsenic-contaminated paddy soils, overexpressing lines contained significantly lower arsenic concentration in rice grain than the wild-type without compromising plant growth or the accumulation of essential nutrients. Overexpression of OsNIP1;1 or OsNIP3;3 provides a route for arsenite to leak out of the stele, thus restricting arsenite loading into the xylem. This strategy is effective in reducing arsenic accumulation in rice grain.

Background Our previous studies concluded that metabolic disorder in the basal portions of rice shoots caused by a lack of cytosolic glutamine synthetase1;2 (GS1;2) resulted in a severe reduction in the outgrowth of tillers. Rice mutants lacking GS1;2 (gs1;2 mutants) showed a remarkable reduction in the contents of both glutamine and asparagine in the basal portions of shoots. In the current study, we attempted to reveal the mechanisms for this decrease in asparagine content using rice mutants lacking either GS1;2 or asparagine synthetase 1 (AS1). The contributions of the availability of glutamine and asparagine to the outgrowth of rice tillers were investigated. Results Rice has two AS genes, and the enzymes catalyse asparagine synthesis from glutamine. In the basal portions of rice shoots, expression of OsAS1, the major species in this tissue, was reduced in gs1;2 mutants, whereas OsAS2 expression was relatively constant. OsAS1 was expressed in phloem companion cells of the nodal vascular anastomoses connected to the axillary bud vasculatures in the basal portions of wild-type shoots, whereas cell-specific expression was markedly reduced in gs1;2 mutants. OsAS1 was up-regulated significantly by NH4+ supply in the wild type but not in gs1;2 mutants. When GS reactions were inhibited by methionine sulfoximine, OsAS1 was up-regulated by glutamine but not by NH4+. The rice mutants lacking AS1 (as1 mutants) showed a decrease in asparagine content in the basal portions of shoots. However, glutamine content and tiller number were less affected by the lack of AS1. Conclusion These results indicate that in phloem companion cells of the nodal vascular anastomoses, asparagine synthesis is largely dependent on glutamine or its related metabolite-responsive AS1. Thus, the decrease in glutamine content caused by a lack of GS1;2 is suggested to result in low expression of OsAS1, decreasing asparagine content. However, the availability of asparagine generated from AS1 reactions is apparently less effective for the outgrowth of tillers. With respect to the tiller number and the contents of glutamine and asparagine in gs1;2 and as1 mutants, the availability of glutamine rather than asparagine in basal portions of rice shoots may be required for the outgrowth of rice tillers. Electronic supplementary material The online version of this article (10.1186/s12284-018-0225-2) contains supplementary material, which is available to authorized users.

Ammonium is a major nitrogen source for plants; it is assimilated into glutamine via a reaction catalyzed by glutamine synthetase (GLN). Arabidopsis expresses four cytosolic GLN genes, GLN1; 1, GLN1; 2, GLN1; 3 and GLN1; 4, in roots. However, the function and organization of these GLN1 isozymes in ammonium assimilation in roots remain unclear. In this study, we aimed to characterize the four GLN1 isozymes. The levels of growth of the wild type and gln1 single and multiple knockout lines were compared in a hydroponic culture at ammonium concentrations of 0.1 and 3 mM. Under the low-ammonium concentration, in single mutants for each GLN1 gene, there was little effect on growth, whereas the triple mutant for GLN1; 1, GLN1; 2 and GLN1; 3 grew slowly and accumulated ammonium. Under the high-ammonium concentration, the single mutant for GLN1; 2 showed 50% decreases in fresh weight and glutamine, whereas the other gln1 single mutants did not show notable changes in the phenotype. The double mutant for GLN1; 1 and GLN1; 2 showed less growth and a lower glutamine concentration than the single mutant for GLN1; 2. Promoter analysis indicated an overlapping expression of GLN1; 1 with GLN1; 2 in the surface layers of the roots. We thus concluded that: (i) at a low concentration, ammonium was assimilated by GLN1; 1, GLN1; 2 and GLN1; 3, and they were redundant; (ii) low-affinity GLN1; 2 could contribute to ammonium assimilation at concentrations ranging from 0.1 to 3 mM; and (iii) GLN1; 1 supported GLN1; 2 within the outer cell layers of the root.

Plant requires nitrogen for the growth, and it use nitrate and ammonium from the environment. Plant suffers from the toxicity when excess ammonium is supplied as a sole nitrogen, although it could be a good nitrogen source for plant growth. We hypothesized that the different responses of ecotypes to ammonium nutrient could partly account for the adaptation of Arabidopsis to an ammonium environment. The purpose of this study is to understand the different responses of ecotypes in ammonium environment. The growth of Arabidopsis thaliana ecotypes, Columbia was compared to those of Arabidopsis thaliana ecotypes, Landsberg erecta in ammonium nutrient. The ratio of shoot dry weight to root dry weight was compared to evaluate the adaptation of two ecotypes. The shoot:root ratio of Landsberg was significantly higher than that of Columbia. T-DNA insertion in cytosolic glutamine synthetase 1;2, one of the essential ammonium assimilatory enzymes, led a decrease of shoot:root ratio. We also measured the isoto...

Nitrogen is an essential element for plant growth. Development of the root system architecture is highly correlated with nitrogen availability from the environment. Recent studies show that auxin response modules are involved in nitrate-dependent lateral root growth. However, the role of auxin in nitrogen metabolism remains to be elucidated. Few researchers have addressed the effect of auxin signaling modules on the use of ammonium for nutrition. The purpose of this study was to describe and examine the relation between auxin signaling modules and ammonium for nutrition. The growth of T-DNA insertion lines for auxin signaling modules was compared with that of a wild type under different nitrogen regimes. Nitrogen use efficiency consists of two components: metabolism and uptake. The nitrogen usage index was calculated following elemental analysis to evaluate nitrogen metabolism. Isotope-labeled ammonium uptake was measured under nitrogen-deficient and -sufficient conditions. Transcriptional levels ...

Root-associated bacterial communities are necessary for healthy plant growth. Nitrate is a signal molecule as well as a major nitrogen source for plant growth. In this study, nitrate-dependent alterations in root-associated bacterial communities and the relationship between nitrate signaling and root-associated bacteria in Arabidopsis were examined. The bacterial community was analyzed by a ribosomal RNA intergenic spacer analysis (RISA) and 16S rRNA amplicon sequencing. The Arabidopsis root-associated bacterial community shifted depending on the nitrate amount and timing of nitrate application. The relative abundance of operational taxonomic units of 25.8% was significantly changed by the amount of nitrate supplied. Moreover, at the family level, the relative abundance of several major root-associated bacteria including Burkholderiaceae, Paenibacillaceae, Bradyrhizobiaceae, and Rhizobiaceae markedly fluctuated with the application of nitrate. These results suggest that the application of nitrate strongly affects root-associated bacterial ecosystems in Arabidopsis. Bulk soil bacterial communities were also affected by the application of nitrate; however, these changes were markedly different from those in root-associated bacteria. These results also suggest that nitrate-dependent alterations in root-associated bacterial communities are mainly affected by plant-derived factors in Arabidopsis. T-DNA insertion plant lines of the genes for two transcription factors involved in nitrate signaling in Arabidopsis roots, NLP7 and TCP20, showed similar nitrate-dependent shifts in root-associated bacterial communities from the wild-type, whereas minor differences were observed in root-associated bacteria. Thus, these results indicate that NLP7 and TCP20 are not major regulators of nitrate-dependent bacterial communities in Arabidopsis roots.

Highlight Temporal and spatial distributions of GLN1;2 and GLN1;3, the two low-affinity cytosolic glutamine synthetase isoforms, determine their contributions to ammonium assimilation in Arabidopsis roots., Glutamine synthetase (GS) catalyzes a reaction that incorporates ammonium into glutamate and yields glutamine in the cytosol and chloroplasts. Although the enzymatic characteristics of the GS1 isozymes are well known, their physiological functions in ammonium assimilation and regulation in roots remain unclear. In this study we show evidence that two cytosolic GS1 isozymes (GLN1;2 and GLN1;3) contribute to ammonium assimilation in Arabidopsis roots. Arabidopsis T-DNA insertion lines for GLN1;2 and GLN1;3 (i.e. gln1;2 and gln1;3 single-mutants), the gln1;2:gln1;3 double-mutant, and the wild-type accession (Col-0) were grown in hydroponic culture with variable concentrations of ammonium to compare their growth, and their content of nitrogen, carbon, ammonium, and amino acids. GLN1;2 and GLN1;3 promoter-dependent green fluorescent protein was observed under conditions with or without ammonium supply. Loss of GLN1;2 caused significant suppression of plant growth and glutamine biosynthesis under ammonium-replete conditions. In contrast, loss of GLN1;3 caused slight defects in growth and Gln biosynthesis that were only visible based on a comparison of the gln1;2 single- and gln1;2:gln1;3 double-mutants. GLN1;2, being the most abundantly expressed GS1 isozyme, markedly increased following ammonium supply and its promoter activity was localized at the cortex and epidermis, while GLN1;3 showed only low expression at the pericycle, suggesting their different physiological contributions to ammonium assimilation in roots. The GLN1;2 promoter-deletion analysis identified regulatory sequences required for controlling ammonium-responsive gene expression of GLN1;2 in Arabidopsis roots. These results shed light on GLN1 isozyme-specific regulatory mechanisms in Arabidopsis that allow adaptation to an ammonium-replete environment.

School non-attendance due to difficulty waking up is a social concern among Japanese students. Many factors result in wake-up difficulty, and it is often difficult to identify the underlying cause at the first visit to a clinic or hospital...

Root-associated bacterial communities are necessary for healthy plant growth. Nitrate is a signal molecule as well as a major nitrogen source for plant growth. In this study, nitrate-dependent alterations in root-associated bacterial communities and the relationship between nitrate signaling and root-associated bacteria in Arabidopsis were examined. The bacterial community was analyzed by a ribosomal RNA intergenic spacer analysis (RISA) and 16S rRNA amplicon sequencing. The Arabidopsis root-associated bacterial community shifted depending on the nitrate amount and timing of nitrate application. The relative abundance of operational taxonomic units of 25.8% was significantly changed by the amount of nitrate supplied. Moreover, at the family level, the relative abundance of several major root-associated bacteria including Burkholderiaceae, Paenibacillaceae, Bradyrhizobiaceae, and Rhizobiaceae markedly fluctuated with the application of nitrate. These results suggest that the application of nitrate strongly affects root-associated bacterial ecosystems in Arabidopsis. Bulk soil bacterial communities were also affected by the application of nitrate; however, these changes were markedly different from those in root-associated bacteria. These results also suggest that nitrate-dependent alterations in root-associated bacterial communities are mainly affected by plant-derived factors in Arabidopsis. T-DNA insertion plant lines of the genes for two transcription factors involved in nitrate signaling in Arabidopsis roots, NLP7 and TCP20, showed similar nitrate-dependent shifts in root-associated bacterial communities from the wild-type, whereas minor differences were observed in root-associated bacteria. Thus, these results indicate that NLP7 and TCP20 are not major regulators of nitrate-dependent bacterial communities in Arabidopsis roots.

Asparagine is synthesized from glutamine by the reaction of asparagine synthetase (AS) and is the major nitrogen form in both xylem and phloem sap in rice (Oryza sativa L.). There are two genes encoding AS, OsAS1 and OsAS2, in rice, but the functions of individual AS isoenzymes are largely unknown. Cell type- and NH4(+)-inducible expression of OsAS1 as well as analyses of knockout mutants were carried out in this study to characterize AS1. OsAS1 was mainly expressed in the roots, with in situ hybridization showing that the corresponding mRNA was specifically accumulated in the three cell layers of the root surface (epidermis, exodermis and sclerenchyma) in an NH4(+)-dependent manner. Conversely, OsAS2 mRNA was abundant in leaf blades and sheathes of rice. Although OsAS2 mRNA was detectable in the roots, its content decreased when NH4(+) was supplied. Retrotransposon-mediated knockout mutants lacking AS1 showed slight stimulation of shoot length and slight reduction in root length at the seedling stage. On the other hand, the mutation caused an approximately 80-90% reduction in free asparagine content in both roots and xylem sap. These results suggest that AS1 is responsible for the synthesis of asparagine in rice roots following the supply of NH4(+). Characteristics of the NH4(+)-dependent increase and the root surface cell-specific expression of OsAS1 gene are very similar to our previous results on cytosolic glutamine synthetase1;2 and NADH-glutamate synthase1 in rice roots. Thus, AS1 is apparently coupled with the primary assimilation of NH4(+) in rice roots.

Plant roots under nitrogen deficient conditions with access to both ammonium and nitrate ions, will take up ammonium first. This preference for ammonium rather than nitrate emphasizes the importance of ammonium assimilation machinery in roots. Glutamine synthetase (GS) and glutamate synthase (GOGAT) catalyze the conversion of ammonium and 2-oxoglutarate to glutamine and glutamate. Higher plants have two GOGAT species, ferredoxin-dependent glutamate synthase (Fd-GOGAT) and nicotinamide adenine dinucleotide (NADH)-GOGAT. While Fd-GOGAT participates in the assimilation of ammonium, which is derived from photorespiration in leaves, NADH-GOGAT is highly expressed in roots and its importance needs to be elucidated. While ammonium as a minor nitrogen form in most soils is directly taken up, nitrate as the major nitrogen source needs to be converted to ammonium prior to uptake. The aim of this study was to investigate and quantify the contribution of NADH-GOGAT to the ammonium assimilation in Arabidopsis (Arabidopsis thaliana Columbia) roots. Quantitative real-time polymerase chain reaction (PCR) and protein gel blot analysis showed an accumulation of NADH-GOGAT in response to ammonium supplied to the roots. In addition the localization of NADH-GOGAT and Fd-GOGAT did not fully overlap. Promoter-β-glucuronidase (GUS) fusion analysis and immunohistochemistry showed that NADH-GOGAT was highly accumulated in non-green tissue like vascular bundles, shoot apical meristem, pollen, stigma and roots. Reverse genetic approaches suggested a reduction in glutamate production and biomass accumulation in NADH-GOGAT transfer DNA (T-DNA) insertion lines under normal CO2 condition. The data emphasize the importance of NADH-GOGAT in the ammonium assimilation in Arabidopsis roots.

This paper has three main aims. First, the paper introduces the concept of integrated reporting ( ) as described by the International Integrated Reporting Council (IIRC). A background to the development of the concept over the 4-year period from the inception of the IIRC in 2010 is provided, culminating in the release by the IIRC of a Consultation Draft (CD) of the framework in March 2013. Second, the paper discusses key issues currently being debated relating to the CD that the IIRC will need to resolve prior to the expected release of their framework in late 2013. This discussion is based on issues identified and reported to the IIRC by a subcommittee of the International Association for Accounting Education and Research (IAAER) comprised of international accounting academics. Finally, the paper identifies a range of potential research issues relating to the development and implementation of .

Rapid eye movement (REM)-related obstructive sleep apnea (OSA) is characterized by apnea and hypopnea events due to airway collapse occurring predominantly or exclusively during REM sleep. Previous studies have reported that REM-related OSA occurs more commonly in women and younger individuals. However, external validity of this tendency has not been confirmed in a large clinical sample. The objective of this study was to evaluate the effect of gender and age on REM-related OSA after adjustment for several covariates based on their established clinical relationships to gender difference in OSA. A total of 3234 Japanese patients with OSA were enrolled in this study. We confirmed that female sex is an important risk factor for REM-related OSA, as reported by previous studies. Moreover, we showed that women aged over 50 years were at a greater risk than those aged under 50 years. These results suggest that hormonal changes in women might play an important role in REM-related OSA and might reflect its unknown pathophysiological characteristics.

Purpose We examined the risk factors for automobile accidents caused by falling asleep while driving in subjects with obstructive sleep apnea syndrome (OSAS). Methods We asked licensed drivers with history of snoring and excessive daytime sleepiness who had undergone polysomnography (PSG) at the Department of Sleep Medicine/Sleep Disorders Center at Aichi Medical University Hospital to complete the questionnaires on accidents caused by falling asleep while driving. As a subjective measure of sleepiness, we used the Epworth sleepiness scale (ESS). Based on PSG results, 2387 subjects diagnosed with OSAS were divided into three groups according to apnea-hypopnea index (AHI): mild-to-moderate (5 ≤ AHI

A spectroscopic method has been applied to measure two-dimensional temperature distribution in luminous spray flames. A test furnace, incorporating a pair of regenerative burners, was used to examine the behavior of heavyfuel oil flames with very high-temperature combustion air. The spectroscopic method used here is based on the two-color thermometry, in which the ratio of spectral intensities of continuum emission is taken at two closely located wavelengths. The two-dimensional distribution of flame temperatures is obtained by the use of a charge-coupled device camera fitted with an optical bandpass filter to provide a much higher resolution compared to that measured with thermocouples. This method was applied to characterize the luminous flames formed under two different conditions during the combustion of heavy oil. The two different kinds of flames formed were either premixed or diffused by the use of air preheats to very high temperatures of 1373 K. A far more uniform temperature distribution was found for the diffusion flame case. In contrast, under premixed conditions, local high-temperature regions were found around the fuel injector exit. The concentration of NO x in flue gas has also been measured for the two different combustion conditions. The temperature distributions obtained from the premixed and diffusion flame conditions are discussed, as are the NO x emission levels from these flames. The results show lower NO x emission for diffusion flames as compared to premixed flames with high air preheats. These results are discussed because standards they provide new for uniform thermal field and ultralow pollution from flames with very high air preheats.

Thermal and chemical characteristics of the flames obtained from an industrial size regenerative combustion furnace have been obtained spectroscopically. The combustion characteristics of diffusion or premixed flames in the regenerative high-temperature air combustion facility have been examined using coal gas as the fuel. The fuel gas composition consisted of H2, hydrocarbon, CO, and N2. Monochromatic images of the flames have been observed in the emission mode using a CCD camera fitted with an optical band pass filter at the desired wavelength. The two-dimensional temperature distribution in the furnace has been determined using the two-line method by utilizing the Swan emission bands from within the flame. The emission intensity profiles of NO, as well as OH and CH radicals have also been observed spectroscopically. The results showed quite uniform two-dimensional temperature distribution and emission intensity of OH and CH radical species for the diffusion flame case as compared to the premixed case using high-temperature combustion air. The premixed flame case showed high local values and large fluctuations in the combustion zone for both emission intensity and temperature distribution. The temperature distribution of soot particles in the premixed flame was also determined using the two-color optical method. The results showed high local value of temperature, similar to that found for the gas temperature using signatures for C2 species at two different wavelengths. In contrast the distribution of temperature for soot particles was different. The location of the maximum soot temperature shifted to downstream positions of the flame as compared to the maximum gas temperature regions measured from the C2 species. The experimental results are discussed in conjunction with those obtained from the heat simulation analyses.

Spontaneous emission spectroscopy has been applied to measure the time-resolved temperature profiles of gaseous fuel flames using high temperature and low oxygen concentration combustion air. Two emission peaks of C2 radical species have been observed at visible wavelengths from propane-air flames. The ratio of these two peaks depends on the flame temperature. The relationship between the ratios of these peaks was correlated with the thermocouple output using a premixed flat flame burner and a multichannel CCD spectrometer. Using this relationship, the flame temperature was determined from the ratio of the C2 peaks. Time-resolved emission intensity profiles of the two C2 bands (two-wavelength image) were observed simultaneously with a high sensitivity video camera fitted with an optical system. The time-resolved temperature profiles were constructed from these intensity profiles by utilizing the previously determined relationship at each pixel. To evaluate fluctuations of flame temperatures, the standard deviation profiles for the temperature profiles have been constructed. This spectroscopic diagnostic technique has been used to measure the profiles of mean flame temperature and temperature fluctuation produced from a concentric diffusion flame using propane as the fuel and high temperature and low oxygen concentration combustion air. In this study, the effect of air-preheat and low oxygen concentration in the combustion air on the subsequent flame temperature and temperature fluctuations has been determined by analyzing the spectra of spontaneous emission from the C2 radicals.

The effect of highly preheated combustion air on the combustion characteristics of propane flames is presented. Data have been obtained on the spatial distribution of emission intensity from selected radicals, fluctuations of emission intensity, and flame temperature using several diagnostic techniques. Several propane and high-temperature air flames, produced with the crossflow diffusion of gas into highly preheated combustion air (obtained by a regenerative combustion facility) having normal oxygen content and reduced oxygen content (

In this study, a spectroscopic method is applied to measure two-dimensional (2D) distributions of flame temperatures during regenerative combustion of natural gas with highly preheated air in a test furnace of industrial size. By using a CCD camera fitted with an optical bandpass filter, the 2D distributions of the continuum emission from soot particles in premix and diffusion flames are measured at two different wavelengths. Subsequently, the temperature is determined on the basis of the two-color 2D thermometry, in which the ratio of the 2D emission intensity distributions is converted into the 2D temperature distribution with much higher spatial resolution in comparison with those obtainable by thermocouples. The emission profiles of NO radiation also were measured and the results are discussed in conjunction with difference in the temperature profiles of the flames.

Higher plants have 2 GOGAT species, Fd-GOGAT and NADH-GOGAT. While Fd-GOGAT mainly assimilates ammonium in leaves, which is derived from photorespiration, the function of NADH-GOGAT, which is highly expressed in roots, (1) needs to be elucidated. The aim of this study was to clarify the role of NADH-GOGAT in Arabidopsis roots. The supply of ammonium to the roots caused an accumulation of NADH-GOGAT, while Fd-GOGAT 1 and Fd-GOGAT 2 showed no response. A promoter-GUS fusion analysis and immunohistochemistry showed that NADH-GOGAT was located in non-green tissues like vascular bundles, shoot apical meristem, pollen, stigma, and roots. The localization of NADH-GOGAT and Fd-GOGAT was not overlapped. NADH-GOGAT T-DNA insertion lines showed a reduction of glutamate and biomass under normal CO2 conditions. These data emphasizes the importance of NADH-GOGAT in the ammonium assimilation of Arabidopsis roots.

This study aims to examine the relationship between corporate characteristics and level of risk reporting and to reveal the impact of issuing regulatory guidelines on risk reporting in annual reports. One hundred non-financial companies were randomly selected from the first section of the Tokyo Stock Exchange. Firstly, we find that, consistent with prior research, the size of the company and the number of risk disclosures are positively correlated. On the other hand, no significant relationship exists between the number of risk disclosures and other corporate characteristics. Secondly, the findings show that most of the companies disclose descriptive risk information but are reluctant to quantify risk. It also shows that the level of risk reporting has increased, at least in quantity, in annual reports after the issuance of regulatory guidelines on risk reporting by the Financial Service Agency.

Rabbit sino-atrial node-right atrium preparations, sympathetic nerve-atria preparations and left atrium preparations were used. N- ethylmaleimide (NEM) in concentrations higher than 10−5 M produced a dose-dependent decrease in atrial rate. At 10−3 M activities the S-A node and right atrium were abolished. The activities were not restored by cysteine and ATP. Transmembrane potentials of S-A nodal pacemaker fibers were affected by 10−3 M NEM. The major changes consisted of a decrease in the slope of diastolic depolarization and an acceleration of repolarization velocity. A tendency for the maximal diastolic potential and the overshoot to decrease were observed. Pacemaker shift frequently occurred. S-A nodal activities were more resistant to NEM than those of the right atrium. Left atrial preparations were driven electrically at different rates. The developed tension-driving rate relationship was not influenced by 10−5 M NEM but was measurably depressed by 10−4 M. The depression of the tension developed at high driving rates (60 to 240 beats/min) was more marked than that at low rates (6 to 30 beats/min). The negative chronotropic response to transmural electrical stimulation applied at the S-A node was slightly reduced in some preparations exposed to 10−7 to 10−5 M NEM, but was blocked by 10−4 M. At the same concentration, slowing of atrial rate produced by acetylcholine was also blocked. The positive chronotropic effect of sympathetic nerve stimulation and of noradrenaline was reduced by 10−4 M NEM. Since the effects of NEM were prevented by preincubation with isomolar concentrations of cysteine, the changes observed were considered to result from an inhibition of the SH groups. Thus, it was concluded that SH groups are involved in the process of contraction and the bioelectrical activity of pacemaker membrane linking automaticity and chronotropic actions of acetylcholine.

Background Asparagine is one of the most dominant organic nitrogen compounds in phloem and xylem sap in a wide range of plant species. Asparaginase (ASNase; EC, 3.5.1.1) catabolizes asparagine into aspartate and ammonium; therefore, it is suggested to play a key role in asparagine metabolism within legume sink organs. However, the metabolic fate of asparagine in source and sink organs during rice seed production remains to be elucidated. Therefore, the main objective of this study is to investigate the asparagine metabolism in a temporal and spatial manner during rice seed production. Results For this purpose, the expression of genes involved in asparagine catabolism, such as asparaginase1 (OsASNase1) and 2 (OsASNase2), were quantitatively measured, and contents of asparagine, aspartate and ammonium ions were determined in sink and source organs during spikelet ripening. Quantitative real-time PCR and in situ localization studies determined that OsASNase2 is expressed in the dorsal vascular bundles and nucellar projection of developing grains, as well as in mesophyll and phloem companion cells of senescent flag leaves. Amino acid measurements revealed that the aspartate concentration is higher than asparagine in both source and sink organs. Conclusion This work suggests that asparaginase dependent asparagine catabolism occurred not only in sink but also in source organs. Electronic supplementary material The online version of this article (doi:10.1186/s12284-017-0143-8) contains supplementary material, which is available to authorized users.