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Abscisic acid (ABA) is a major phytohormone that regulates abiotic stress responses and development. SNF1-rerated protein kinase 2 (SnRK2) is a key regulator of ABA signaling. To isolate compounds which directly affect SnRK2 activity, we optimized a fluorescence-based system for high-throughput screening (HTS) of SnRK2 kinase regulators. Using this system, we screened a chemical library consisting of 16,000 compounds and identified ten compounds (INH1-10) as potential SnRK2 inhibitors. Further characterization of these compounds by in vitro phosphorylation assays confirmed that three of the ten compounds were SnRK2-specific kinase inhibitors. In contrast, seven of ten compounds inhibited ABA-responsive gene expression in Arabidopsis cells. From these results, INH1 was identified as a SnRK2-specific inhibitor in vitro and in vivo. We propose that INH1 could be a lead compound of chemical tools for studying ABA responses in various plant species., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The regional budget of methane (CH 4 ) emissions for East Asia, a crucial region in the global greenhouse gas budget, was quantified for 1990-2015 with a bottom-up method based on inventories and emission model simulations. Anthropogenic emissions associated with fossil fuel extraction, industrial activities, waste management, and agricultural activities were derived from the Emission Database for Global Atmospheric Research version 4.3.2 and compared with other inventories. Emissions from natural wetlands and CH 4 uptake by upland soil oxidation were estimated using the Vegetation Integrative SImulator for Trace gases (VISIT), a biogeochemical model that considers historical land use and climatic conditions. Emissions from biomass burning and termites were calculated using satellite and land-use data combined with empirical emission factors. The resulting average annual estimated CH 4 budget for 2000-2012 indicated that East Asia was a net source of 67.3 Tg CH 4 yr -1 , of which 88.8% was associated with anthropogenic emissions. The uncertainty (±standard deviation) of this estimate, ±14 Tg CH 4 yr -1 , stemmed from data and model inconsistencies. The increase of the net flux from 60.2 Tg CH 4 yr -1 in 1990 to 78.0 Tg CH 4 yr -1 in 2012 was due mainly to increased emissions by the fossil fuel extraction and livestock sectors. Our results showed that CH 4 was a crucial component of the regional greenhouse gas budget. A spatial analysis using 0.25° × 0.25° grid cells revealed emission hotspots in urban areas, agricultural areas, and wetlands. These hotspots were surrounded by weak sinks in upland areas. The estimated natural and anthropogenic emissions fell within the range of independent estimates, including top-down estimates from atmospheric inversion models. Such a regional accounting is an effective way to elucidate climatic forcings and to develop mitigation policies. Further studies, however, are required to reduce the uncertainties in the budget., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

We investigated the effects of lignins as diet components on the physiological activities of a lower termite, Coptotermes formosanus Shiraki. Artificial diets composed of polysaccharides with and without purified lignins (milled-wood lignins) from Japanese cedar (softwood), Japanese beech (hardwood), and rice (grass), were fed to C. formosanus workers. The survival and body mass of the workers as well as the presence of three symbiotic protists in the hindguts of the workers were then periodically examined. The survival rates of workers fed on diets containing lignins were, regardless of the lignocellulose diet sources, significantly higher than those of workers fed on only polysaccharides. In addition, it was clearly observed that all the tested lignins have positive effects on the maintenance of two major protists in the hindguts of C. formosanus workers, i.e., Pseudotrichonympha grassii and Holomastigotoides hartmanni. Overall, our data suggest that the presence of lignin is crucial to maintaining the physiological activities of C. formosanus workers during their lignocellulose decomposition. Our data also suggested that some components, possibly minerals and/or non-structural carbohydrates, in grass lignocellulose negatively affect the survival of C. formosanus workers as well as the present rate of the symbiotic protists in their hindguts., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background: Adaptive statistical iterative reconstruction (ASIR) has been used to reduce radiation dose in cardiac computed tomography. However, change of image parameters by ASIR as compared to filtered back projection (FBP) may influence quantification of coronary calcium., Objective: To investigate the influence of ASIR on calcium quantification in comparison to FBP., Methods: In 352 patients, CT images were reconstructed using FBP alone, FBP combined with ASIR 30%, 50%, 70%, and ASIR 100% based on the same raw data. Image noise, plaque density, Agatston scores and calcium volumes were compared among the techniques., Results: Image noise, Agatston score, and calcium volume decreased significantly with ASIR compared to FBP (each P < 0.001). Use of ASIR reduced Agatston score by 10.5% to 31.0%. In calcified plaques both of patients and a phantom, ASIR decreased maximum CT values and calcified plaque size., Conclusion: In comparison to FBP, adaptive statistical iterative reconstruction (ASIR) may significantly decrease Agatston scores and calcium volumes., (Copyright © 2016 Society of Cardiovascular Computed Tomography. Published by Elsevier Inc. All rights reserved.)

Eugenol, a volatile phenylpropene found in many plant species, exhibits antibacterial and acaricidal activities. This study attempted to modify the production of eugenol and its glycosides by introducing petunia coniferyl alcohol acetyltransferase (PhCFAT) and eugenol synthase (PhEGS) into hybrid aspen. Gas chromatography analyses revealed that wild-type hybrid aspen produced small amount of eugenol in leaves. The heterologous overexpression of PhCFAT alone resulted in up to 7-fold higher eugenol levels and up to 22-fold eugenol glycoside levels in leaves of transgenic aspen plants. The overexpression of PhEGS alone resulted in a subtle increase in either eugenol or eugenol glycosides, and the overexpression of both PhCFAT and PhEGS resulted in significant increases in the levels of both eugenol and eugenol glycosides which were nonetheless lower than the increases seen with overexpression of PhCFAT alone. On the other hand, overexpression of PhCFAT in transgenic Arabidopsis and tobacco did not cause any synthesis of eugenol. These results indicate that aspen leaves, but not Arabidopsis and tobacco leaves, have a partially active pathway to eugenol that is limited by the level of CFAT activity and thus the flux of this pathway can be increased by the introduction of a single heterologous gene., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The ability of plants to tolerate drought conditions is crucial for agricultural production worldwide. Recent progress has been made in our understanding of gene expression, transcriptional regulation and signal transduction in plant responses to drought. Molecular and genomic analyses have facilitated gene discovery and enabled genetic engineering using several functional or regulatory genes to activate specific or broad pathways related to drought tolerance in plants. Several lines of evidence have indicated that molecular tailoring of genes has the potential to overcome a number of limitations in creating drought-tolerant transgenic plants. Recent studies have increased our understanding of the regulatory networks controlling the drought stress response and have led to practical approaches for engineering drought tolerance in plants.

Clonidine, an alpha(2)-adrenoceptor agonist, has been reported to inhibit gastric and small intestinal motility in rats. Whether clonidine also inhibits colonic motility is still not clear. The aim of this study was to examine the effect of clonidine on colonic motility and its possible site of action in adult Wistar rats. Colonic motilities in anesthetized rats in vivo or motilities of the isolated colon of rats in vitro were recorded. Clonidine was administered intravenously (i.v.) and intracerebroventricularly (i.c.v.) in vivo while bath administration was used in in vitro study. Clonidine i.v. or i.c.v. significantly inhibited colonic motility. This inhibitory effect was antagonized by pre-administration of yohimbine, an alpha(2)-adrenoceptor antagonist, but not by pre-administration of prazosin, an alpha(1)-adrenoceptor antagonist. Also, we have unpublished data indicating that the sympathectomy antagonized the inhibitory effect of systemically administered clonidine. A significant depression of colonic motility on the isolated colon was induced by bath administration of noradrenaline, while no such inhibition was seen by clonidine. The results of the present study suggested that clonidine inhibits colonic motility in rats through activation of central alpha(2)-adrenergic receptor.

The role of the sympathetic nervous system in the inhibitory effect of morphine on colonic motility was investigated in male adult Wistar rats. The responses of colonic motility and blood pressure to the intravenous administration of morphine under urethane anesthesia were recorded. Sympathectomy (6-hydroxydopamine) or pretreatment with phentolamine, an alpha-adrenoreceptor antagonist (3.15 microM/kg, i.v.), or propranolol, a beta-adrenoreceptor antagonist (3.38 microM/kg, i.v.), on the inhibitory effect of intravenously administered morphine on colonic motility were observed. The results of the investigation showed that a significant depression of colonic motility occurred in untreated rats following intravenous administration of morphine, while no significant effect was seen in the sympathectomized. Pretreatment with phentolamine or propranolol also significantly attenuated the depression of colonic motility induced by morphine. Morphine also produced a transient depression of the blood pressure. However, this inhibitory effect of morphine on blood pressure was intensified after sympathectomy or pretreatment with either phentolamine or propranolol. We conclude that sympathetic activity plays an important role in the inhibitory effects of morphine on colonic motility and that both alpha- and beta-adrenoreceptors are involved.

Cinnamyl alcohol dehydrogenase (CAD) (EC 1.1.1.195) from a dicot, Aralia cordata, was purified to homogeneity and its properties were characterized. The enzyme shows a preference for cinnamyl alcohols and cinnamyl aldehydes as substrates. The M(r) is estimated at 72,000. The enzyme is composed of two heterogeneous subunits of slightly different sizes, and it differs from the bean enzyme in the size of subunits. Partial amino acid sequencing of the purified enzyme was carried out both from the N-terminus and using selected peptides obtained by cyanogen bromide cleavage.

A crude cell-free extract from Forsythia intermedia catalyses the formation of (-)secoisolariciresinol, and not its (+) enantiomer, when incubated with coniferyl alcohol in the presence of NAD(P)H and H2O2. This is the first report of an enzyme reaction specifically involved in the coupling reaction between two phenylpropanoid monomers to afford an optically pure lignan.

The catalytic C-C bond cleavage of a lignin model compound was investigated by use of tetraphenylporphyrinatoiron(III)chloride as a model for enzymic degradation of lignin. The C-C bond of the lignin model compound 1,2-bis(4-ethoxy-3-methoxyphenyl) propane-1,3-diol was oxidatively cleaved by catalysis of iron-porphyrins in the presence of tert-butylhydroperoxide or iodosylbenzene at a room temperature. The products formed after complete oxidation of the substrate were identified as 4-O-ethylvanillin, alpha-hydroxy-4-ethoxy-3-methoxyacetophenone, 4-O-ethylvanillic acid, 4-ethoxy-3-methoxyphenylglycol, 4-ethoxy-3-methoxy-alpha-(4-ethoxy-3-methoxyphenyl)-beta-hydroxypropi ophenone and formaldehyde.

Phenolic beta-1 lignin substructure model compounds, 1-(3,5-dimethoxy-4-hydroxy-phenyl)-2-(3,5-dimethoxy-4-ethoxyphenyl)propa ne-1, 3-diol (I) and 1-(3,5-dimethoxy-4-ethoxyphenyl)-2-(3, 5-dimethoxy-4-hydroxyphenyl)propane-1,3-diol (II) were degraded by laccase of Coriolus versicolor. Substrate I was converted to 1-(3,5-dimethoxy-4-hydroxyphenyl)-2-(3,5-dimethoxy-4-ethoxyphenyl)-3- hydroxypropanone (III), 1-(3,5-dimethoxy-4-ethoxyphenyl)-2-hydroxyethanone (IV), syringaldehyde (V), 1-(3,5-dimethoxy-4-ethoxyphenyl)-3-hydroxypropanal (VI), 2,6-dimethoxy-p-hydroquinone (VII), and 2,6-dimethoxy-p-benzoquinone (VIII). Furthermore, incorporations of 18O of 18O2 into ethanone (IV) and 18O of H218O into hydroquinone (VII) and benzoquinone (VIII) were confirmed. Substrate II gave 1-(3,5-dimethoxy-4-hydroxyphenyl)ethane-1, 2-diol (IX), 1-(3,5-dimethoxy-4-hydroxyphenyl)-2-hydroxyethanone (X), and 3,5-dimethoxy-4-ethoxybenzaldehyde (XI). Also 18O of H218O was incorporated into glycol (IX) and ethanone (X). Based on the structures of the degradation products and the isotopic experiments, it was established that three types of reactions occurred via phenoxy radicals of substrates caused by laccase: (i) C alpha-C beta cleavage (between C1 and C2 carbons); (ii) alkyl-aryl cleavage (between C1 carbon and aryl group); and (iii) C alpha (C1) oxidation.